Solid forms of isoquinolinones, and process of making, composition comprising, and methods of using the same

ABSTRACT

Solid forms of chemical compounds that modulate kinase activity, including PI3 kinase activity, and compounds, pharmaceutical compositions, and methods of treatment of diseases and conditions associated with kinase activity, including PI3 kinase activity, are described herein. Also provided herein are processes for preparing compounds, polymorphic forms, cocrystals, and amorphous forms thereof, and pharmaceutical compositions thereof.

This application claims priority to U.S. Provisional Application Nos.62/218,486, filed Sep. 14, 2015, and 62/218,493, filed Sep. 14, 2015,the entireties of which are incorporated herein by reference.

1. BACKGROUND

The activity of cells can be regulated by external signals thatstimulate or inhibit intracellular events. The process by whichstimulatory or inhibitory signals are transmitted into and within a cellto elicit an intracellular response is referred to as signaltransduction. Over the past decades, cascades of signal transductionevents have been elucidated and found to play a central role in avariety of biological responses. Defects in various components of signaltransduction pathways have been found to account for a vast number ofdiseases, including numerous forms of cancer, inflammatory disorders,metabolic disorders, vascular and neuronal diseases (Gaestel et al.Current Medicinal Chemistry (2007) 14:2214-2234).

Kinases represent a class of important signaling molecules. Kinases cangenerally be classified into protein kinases and lipid kinases, andcertain kinases exhibit dual specificities. Protein kinases are enzymesthat phosphorylate other proteins and/or themselves (i.e.,autophosphorylation). Protein kinases can be generally classified intothree major groups based upon their substrate utilization: tyrosinekinases which predominantly phosphorylate substrates on tyrosineresidues (e.g., erb2, PDGF receptor, EGF receptor, VEGF receptor, src,abl), serine/threonine kinases which predominantly phosphorylatesubstrates on serine and/or threonine residues (e.g., mTorC1, mTorC2,ATM, ATR, DNA-PK, Akt), and dual-specificity kinases which phosphorylatesubstrates on tyrosine, serine and/or threonine residues.

Lipid kinases are enzymes that catalyze the phosphorylation of lipids.These enzymes, and the resulting phosphorylated lipids and lipid-derivedbiologically active organic molecules play a role in many differentphysiological processes, including cell proliferation, migration,adhesion, and differentiation. Certain lipid kinases are membraneassociated and they catalyze the phosphorylation of lipids contained inor associated with cell membranes. Examples of such enzymes includephosphoinositide(s) kinases (e.g., PI3-kinases, PI4-Kinases),diacylglycerol kinases, and sphingosine kinases.

Phosphoinositide 3-kinases (PI3Ks) constitute a unique and conservedfamily of intracellular lipid kinases that phosphorylate the 3′-OH groupon phosphatidylinositols or phosphoinositides. The PI3K family comprises15 kinases with distinct substrate specificities, expression patterns,and modes of regulation. The class I PI3Ks (p110α, p110β, p110δ, andp110γ) are typically activated by tyrosine kinases or G-protein coupledreceptors to generate a lipid product termed PIP₃, which engagesdownstream effectors such as those in the Akt/PDK1 pathway, mTOR, theTec family kinases, and the Rho family GTPases. The class II and IIIPI3Ks play a key role in intracellular trafficking through the synthesisof PI(3)P and PI(3,4)P2.

The PI3K signaling pathway is one of the most highly mutated systems inhuman cancers. PI3K signaling is also a key factor in many otherdiseases in humans. PI3K signaling is involved in many disease statesincluding allergic contact dermatitis, rheumatoid arthritis,osteoarthritis, inflammatory bowel diseases, chronic obstructivepulmonary disorder, psoriasis, multiple sclerosis, asthma, disordersrelated to diabetic complications, and inflammatory complications of thecardiovascular system such as acute coronary syndrome.

Many inhibitors of PI3Ks have been generated. While such compounds areoften initially evaluated for their activity when dissolved in solution,solid state characteristics such as polymorphism play an important role.Polymorphic forms of a drug substance, such as an inhibitor of PI3K, canhave different chemical and physical properties, includingcrystallinity, melting point, chemical reactivity, solubility,dissolution rate, optical and mechanical properties, vapor pressure, anddensity. These properties can have a direct effect on the ability toprocess or manufacture a drug substance and the drug product. Moreover,polymorphism is often a factor under regulatory review of the “sameness”of drug products from various manufacturers. For example, polymorphismhas been evaluated in compounds such as warfarin sodium, famotidine, andranitidine. Polymorphism can affect the quality, safety, and/or efficacyof a drug product, such as a kinase inhibitor. Thus, research directedtowards polymorphs of PI3K inhibitors and processes for preparingpolymorphs of PI3K inhibitors represents a significantly useful field ofinvestigation in the development of active pharmaceutical ingredients(APIs).

In addition, PI3K inhibitors have been used to treat various diseasesand disorders in humans (e.g., in clinical trials). For the productionof a drug substance intended for use in humans, current GoodManufacturing Practices (GMP) are applicable. Procedures need to be inplace that can control the levels of impurities and ensure that APIproducts are produced, which consistently meet their predeterminedspecifications. Thus, a significant need exists for a process to preparePI3K inhibitors suitable for human use, particularly on a commercialscale, that is, inter alia, safe, scalable, efficient, economicallyviable, and/or having other desirable properties. Among other entities,disclosed herein are polymorphic forms of PI3K inhibitors which addressthese needs and provide exemplary advantages.

2. SUMMARY

Provided herein are solid forms comprising a compound of formula (I)(also referred as Compound 1 herein):

or a salt, or solvate (e.g., hydrate), or solvate of a salt thereof, ora mixture thereof. Also provided herein are methods of synthesizing thesolid forms.

The solid forms provided herein include, but are not limited to,hydrates, anhydrates, solvates of Compound 1 and salts and cocrystalsthereof. The solid forms provided herein are useful as activepharmaceutical ingredients for the preparation of formulations for usein animals or humans. Thus, embodiments herein encompass the use ofthese solid forms as a final drug product. Certain embodiments providesolid forms useful in making final dosage forms with improvedproperties, e.g., powder flow properties, compaction properties,tableting properties, stability properties, and excipient compatibilityproperties, among others, that are needed for manufacturing, processing,formulation and/or storage of final drug products. Certain embodimentsherein provide pharmaceutical compositions comprising a single-componentcrystal form, and/or a multiple-component crystal form comprising thecompound of formula (I) and a pharmaceutically acceptable diluent,excipient or carrier.

In one embodiment, the solid form is a crystalline form. In oneembodiment, the solid form further comprises a coformer. In oneembodiment, the solid form comprising Compound 1 and a coformer is acocrystal. In another embodiment, the solid form is an amorphous form.

Also provided herein are pharmaceutical compositions, single unit dosageforms, dosing regimens and kits comprising the amorphous form providedherein.

Also provided herein are methods for treating, preventing, and managingvarious disorders using the compositions and amorphous form providedherein. The methods comprise administering to a patient in need of suchtreatment or management a therapeutically effective amount of a compoundprovided herein. Further provided are methods of preventing variousdiseases and disorders, which comprise administering to a patient inneed of such prevention a prophylactically effective amount of acompound provided herein.

Further provided herein are processes of preparing a compound of formula(I), or a salt, or solvate (e.g., hydrate), or solvate of a saltthereof, or a mixture thereof.

Further provided herein are methods for analyzing a material for thepresence or amount of a solid form provided herein, comprising providinga material comprising a compound of formula (I), or a salt, solvate(e.g., hydrate), or solvate of a salt thereof, or a mixture thereof; andusing a characterization method to determine whether a signatorycharacteristic associated with the solid form is present in the materialby comparing the characteristic obtained from the material with areference signatory characteristic; wherein the existence of acharacteristic substantially identical to the reference signatorycharacteristic indicates the presence of the solid form in the material.

3. INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference in their entiretiesand to the same extent as if each individual publication, patent, orpatent application was specifically and individually indicated to beincorporated by reference.

4. BRIEF DESCRIPTION OF FIGURES

FIG. 1 is a representative X-ray powder diffraction (XRPD) pattern ofForm 1 of Compound 1.

FIG. 2 is a representative overlay of thermal gravimetric analysis (TGA)and differential scanning calorimetry (DSC) thermograms for Form 1 ofCompound 1.

FIG. 3 is a representative gravimetric vapour sorption (GVS) isothermplot of Form 1 of Compound 1.

FIG. 4 is a representative XRPD pattern of Form 2 of Compound 1.

FIG. 5 is a representative overlay of TGA and DSC thermograms for Form 2of Compound 1.

FIG. 6 is a representative GVS isotherm plot of Form 2 of Compound 1.

FIG. 7 is another representative overlay of TGA and DSC thermograms forForm 2 of Compound 1.

FIG. 8 is another representative XRPD pattern of form 2 of Compound 1.

FIG. 9 is a representative XRPD pattern of Form 3 of Compound 1.

FIG. 10 is a representative overlay of TGA and DSC thermograms for Form3 of Compound 1.

FIG. 11 is a representative XRPD pattern of Form 4 of Compound 1.

FIG. 12 is a representative overlay of TGA and DSC thermograms for Form4 of Compound 1.

FIG. 13 is a representative XRPD pattern of Form 5 of Compound 1.

FIG. 14 is a representative overlay of TGA and DSC thermograms for Form5 of Compound 1.

FIG. 15 is a representative XRPD pattern of Form 6 of Compound 1.

FIG. 16 is a representative overlay of TGA and DSC thermograms for Form6 of Compound 1.

FIG. 17 is a representative GVS isotherm plot of Form 6 of Compound 1.

FIG. 18 is a representative XRPD pattern of Form 7 of Compound 1.

FIG. 19 is a representative overlay of TGA and DSC thermograms for Form7 of Compound 1.

FIG. 20 is a representative XRPD pattern of Form 8 of Compound 1.

FIG. 21 is a representative ORTEP plot of Form 2 of Compound 1.

FIG. 22 is a representative XRPD pattern of Form P1C3 of a cocrystal ofCompound 1 and L-tartaric acid.

FIG. 23 is a representative TGA and DSC analysis of Form P1C3 of acocrystal of Compound 1 and L-tartaric acid.

FIG. 24 is a representative XRPD of Form P1C9 of a cocrystal of Compound1 and salicylic acid.

FIG. 25 is a representative TGA vs. DSC analysis of Form P1C9 of acocrystal of Compound 1 and salicylic acid.

FIG. 26 is a representative XRPD analysis of Form P2C9 of a cocrystal ofCompound 1 and salicylic acid.

FIG. 27 is a representative TGA and DSC analysis of Form P2C9 of acocrystal of Compound 1 and salicylic acid.

FIG. 28 is a representative GVS analysis of Form P1C3 of a cocrystal ofCompound 1 and L-tartaric acid.

FIG. 29 shows Compound 1 solubility in ethanol/water and correspondingethanol content in isolated Compound 1.

FIG. 30 is a representative crystal structure of Form 1 of Compound 1obtained by heating 13.5 mg/ml Compound 1 in 80% ethanol/water to 60° C.then cool to room temperature.

FIG. 31 is a representative TGA analysis of spray dried Compound 1.

FIG. 32 is a representative TGA analysis of spray dried Compound 1 andPVP/VA 64.

FIG. 33 is a representative TGA analysis of spray dried Compound 1 andHPMC-AS.

FIG. 34A shows the conversion at 2 hours (% area) for the couplingreaction of 4-iodo-1-methyl-1H-pyrazole with trimethylsilylacetylene;FIG. 34B shows the reaction conversion at 24 hours (% area); FIG. 34Cshows the product/diyne ratio (% area) at 24 hours; and FIG. 34D showsthe diyne content at 24 hours (% area).

5. DETAILED DESCRIPTION

5.1 Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart.

As used in the specification and claims, the singular form “a”, “an” and“the” includes plural references unless the context clearly dictatesotherwise.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. As used herein, the terms “about”and “approximately” when used in combination with a numeric value orrange of values mean that the value or range of values may deviate to anextent deemed reasonable to one of ordinary skill in the art, e.g.,within experimental variability (or within statistical experimentalerror), and thus the numeric value or range of values can vary from, forexample, between 1% and 15%, between 1% and 10%, between 1% and 5%,between 0.5% and 5%, and between 0.5% and 1%, of the stated numericvalue or range of values. As disclosed herein, every instance where anumeric value or range of values preceded by the term “about” alsoincludes the embodiment of the given value(s). For example, “about 3°C.” discloses the embodiment of the temperature being “3° C.”. The terms“about” and “approximately” are used completely interchangeablethroughout the disclosure. The term “between” includes the endpointnumbers on both limits of the range. For example, the range described by“between 3 and 5” is inclusive of the numbers “3” and “5”. As usedherein, a tilde (i.e., “˜”) preceding a numerical value or range ofvalues indicates “about” or “approximately.”

As used herein, and unless otherwise specified, “agent” or “biologicallyactive agent” or “second active agent” refers to a biological,pharmaceutical, or chemical compound or other moiety. Non-limitingexamples include simple or complex organic or inorganic molecules, apeptide, a protein, an oligonucleotide, an antibody, an antibodyderivative, antibody fragment, a vitamin derivative, a carbohydrate, atoxin, or a chemotherapeutic compound. Various compounds can besynthesized, for example, small molecules and oligomers (e.g.,oligopeptides and oligonucleotides), and synthetic organic compoundsbased on various core structures. In addition, various natural sourcescan provide compounds for screening, such as plant or animal extracts,and the like. A skilled artisan can readily recognize that there is nolimit as to the structural nature of the agents of the presentdisclosure.

As used herein, and unless otherwise specified, the term “agonist”refers to a compound having the ability to initiate or enhance abiological function of a target protein, whether by enhancing orinitiating the activity or expression of the target protein.Accordingly, the term “agonist” is defined in the context of thebiological role of the target protein. While agonists provided hereincan specifically interact with (e.g., bind to) the target, compoundsthat initiate or enhance a biological activity of the target protein byinteracting with other members of the signal transduction pathway ofwhich the target protein is a member are also specifically includedwithin this definition.

As used herein, and unless otherwise specified, the terms “antagonist”and “inhibitor” are used interchangeably, and they refer to a compoundhaving the ability to inhibit a biological function of a target protein,whether by inhibiting the activity or expression of the target protein.Accordingly, the terms “antagonist” and “inhibitors” are defined in thecontext of the biological role of the target protein. While antagonistsprovided herein can specifically interact with (e.g., bind to) thetarget, compounds that inhibit a biological activity of the targetprotein by interacting with other members of the signal transductionpathway of which the target protein is a member are also specificallyincluded within this definition. In one embodiment, a biologicalactivity inhibited by an antagonist is associated with the development,growth, or spread of a tumor, or an undesired immune response, e.g., asmanifested in autoimmune disease.

As used herein, and unless otherwise specified, an “anti-cancer agent”,“anti-tumor agent” or “chemotherapeutic agent” refers to any agentuseful in the treatment of a neoplastic condition. One class ofanti-cancer agents comprises chemotherapeutic agents. As used herein,and unless otherwise specified, “chemotherapy” means the administrationof one or more chemotherapeutic drugs and/or other agents to a cancerpatient by various methods, including intravenous, oral, intramuscular,intraperitoneal, intravesical, subcutaneous, transdermal, buccal, orinhalation or in the form of a suppository.

As used herein, and unless otherwise specified, the term “cellproliferation” refers to a phenomenon by which the cell number haschanged as a result of division. In one embodiment, this term alsoencompasses cell growth by which the cell morphology has changed (e.g.,increased in size) consistent with a proliferative signal.

As used herein, and unless otherwise specified, the term“co-administration,” “administered in combination with,” and theirgrammatical equivalents, encompasses administration of two or moreagents to an animal either simultaneously or sequentially. In oneembodiment, both agents and/or their metabolites are present in theanimal at the same time. In one embodiment, co-administration includessimultaneous administration in separate compositions, administration atdifferent times in separate compositions, or administration in acomposition in which both agents are present.

As used herein, and unless otherwise specified, the term “effectiveamount” or “therapeutically effective amount” refers to an amount of acompound described herein that is sufficient to effect an intendedapplication or effect, including, but not limited to, disease treatment,as defined herein. The therapeutically effective amount can varydepending upon the intended application (in vitro or in vivo), or thesubject and disease condition being treated, e.g., the weight and age ofthe subject, the severity of the disease condition, the manner ofadministration, and the like, which can be determined by one of ordinaryskill in the art. The term can also apply to a dose that will induce aparticular response in target cells, e.g., reduction of plateletadhesion and/or cell migration. The specific dose will vary depending onthe particular compounds chosen, the dosing regimen to be followed,whether it is administered in combination with other compounds, timingof administration, the tissue to which it is administered, and thephysical delivery system in which it is carried.

As used herein, and unless otherwise specified, the terms “treatment”,“treating”, “palliating” and “ameliorating” are used interchangeablyherein, and refer to an approach for obtaining beneficial or desiredresults, including, but not limited to, a therapeutic benefit. In oneembodiment, therapeutic benefit means eradication or amelioration of theunderlying disorder being treated. In one embodiment, a therapeuticbenefit is achieved with the eradication or amelioration of one or moreof the physiological symptoms associated with the underlying disorder,such that an improvement is observed in the patient, notwithstandingthat the patient can still be afflicted with the underlying disorder.

As used herein, and unless otherwise specified, the terms “prevention”and “preventing” refer to an approach for obtaining beneficial ordesired results including, but not limited to, prophylactic benefit. Inone embodiment, prophylactic benefit includes delaying or eliminatingthe appearance of a disease or condition, delaying or eliminating theonset of symptoms of a disease or condition, slowing, halting, orreversing the progression of a disease or condition, or any combinationthereof. To obtain prophylactic benefit, the compositions can beadministered to a patient at risk of developing a particular disease, orto a patient reporting one or more of the physiological symptoms of adisease, even though a diagnosis of this disease can or cannot have beenmade.

As used herein, and unless otherwise specified, “signal transduction” isa process during which stimulatory or inhibitory signals are transmittedinto and within a cell to elicit an intracellular response. A modulatorof a signal transduction pathway refers to a compound which modulatesthe activity of one or more cellular proteins mapped to the samespecific signal transduction pathway. A modulator can augment (agonist)or suppress (antagonist) the activity of a signaling molecule.

As used herein, and unless otherwise specified, the term “selectiveinhibition” or “selectively inhibit” as applied to a biologically activeagent refers to the agent's ability to selectively reduce the targetsignaling activity as compared to off-target signaling activity, viadirect or interact interaction with the target.

As used herein, and unless otherwise specified, the term “in vivo”refers to an event that takes place in a subject's body.

As used herein, and unless otherwise specified, the term “in vitro”refers to an event that takes places outside of a subject's body. Forexample, an in vitro assay encompasses any assay run outside of asubject assay. In vitro assays encompass cell-based assays in whichcells alive or dead are employed. In one embodiment, in vitro assaysalso encompass a cell-free assay in which no intact cells are employed.

“Subject” to which administration is contemplated includes, but is notlimited to, humans (i.e., a male or female of any age group, e.g., apediatric subject (e g., infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or otherprimates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, includingcommercially relevant mammals such as cattle, pigs, horses, sheep,goats, cats, and/or dogs; and/or birds, including commercially relevantbirds such as chickens, ducks, geese, quail, and/or turkeys.

As used herein, and unless otherwise specified, “radiation therapy”means exposing a patient, using routine methods and compositions knownto the practitioner, to radiation emitters such as alpha-particleemitting radionuclides (e.g., actinium and thorium radionuclides), lowlinear energy transfer (LET) radiation emitters (e.g., beta emitters),conversion electron emitters (e.g., strontium-89 andsamarium-153-EDTMP), or high-energy radiation, including withoutlimitation, x-rays, gamma rays, and neutrons.

As used herein, the term “combining” refers to bringing one or morechemical entities into association with another one or more chemicalentities. Combining includes the processes of adding one or morecompounds to a solid, liquid or gaseous mixture of one or more compounds(the same or other chemical entities), or a liquid solution ormultiphasic liquid mixture. The act of combining includes the process orprocesses of one or more compounds reacting (e.g., bond formation orcleavage; salt formation, solvate formation, chelation, or othernon-bond altering association) with one or more compounds (the same orother chemical entities). The act of combining can include alteration ofone or more compounds, such as by isomerization (e.g., tautomerization,resolution of one isomer from another, or racemization).

As used herein, and unless otherwise specified, a “one-pot” processrefers to a process of preparing a desired product, wherein allreactants are added simultaneously or successively, and wherein noseparation, isolation, and/or purification of any intermediate formed isconducted before the formation of the desired product is substantiallycomplete. A “one-pot” process is preferably conducted in a singlecontainer, but may be conducted in more than one container.

As used herein, the term “recovering” includes, but is not limited to,the action of obtaining one or more compounds by collection duringand/or after a process step as disclosed herein, and the action ofobtaining one or more compounds by separation of one or more compoundsfrom one or more other chemical entities during and/or after a processstep as disclosed herein. The term “collection” refers to any action(s)known in the art for this purpose, including, but not limited to,filtration, decanting a mother liquor from a solid to obtain one or morecompounds, and evaporation of liquid media in a solution or othermixture to afford a solid, oil, or other residue that includes one ormore compounds. The solid can be crystalline, acrystalline, partiallycrystalline, amorphous, containing one or more polymorphs, a powder,granular, of varying particle sizes, of uniform particle size, amongother characteristics known in the art. An oil can vary in color andviscosity, and include one or more solid forms as a heterogeneousmixture, among other characteristics known in the art. The term“separation” refers to any action(s) known in the art for this purpose,including, but not limited to, isolating one or more compounds from asolution or mixture using, for example, seeded or seedlesscrystallization or other precipitation techniques (e.g., adding ananti-solvent to a solution to induce compound precipitation; heating asolution, then cooling to induce compound precipitation; scratching thesurface of a solution with an implement to induce compoundprecipitation), and distillation techniques. Recovering one or morecompounds can involve preparation of a salt, solvate, hydrate, chelateor other complexes of the same, then collecting or separating asdescribed above.

As used herein, a “pharmaceutically acceptable form” of a disclosedFormula (I) includes, but is not limited to, pharmaceutically acceptablesalts, hydrates, solvates, chelates, non-covalent complexes, isomers,prodrugs, and isotopically labeled derivatives thereof, and mixturesthereof. Hence, the terms “chemical entity” and “chemical entities” alsoencompass pharmaceutically acceptable salts, hydrates, solvates,chelates, non-covalent complexes, isomers, prodrugs, and isotopicallylabeled derivatives, and mixtures thereof. In some embodiments, apharmaceutically acceptable form of a disclosed Formula (I) includes asalt, a solvate, or a hydrate thereof.

In certain embodiments, the pharmaceutically acceptable form is apharmaceutically acceptable salt. As used herein, the term“pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of subjects without undue toxicity, irritation,allergic response and the like, and are commensurate with a reasonablebenefit/risk ratio. Pharmaceutically acceptable salts are well known inthe art. For example, Berge et al. describes pharmaceutically acceptablesalts in detail in J. Pharmaceutical Sciences (1977) 66:1-19.Pharmaceutically acceptable salts of the compounds provided hereininclude those derived from suitable inorganic and organic acids andbases. Inorganic acids from which salts can be derived include, but arenot limited to, hydrochloric acid, hydrobromic acid, sulfuric acid,nitric acid, phosphoric acid, and the like. Organic acids from whichsalts can be derived include, but are not limited to, acetic acid,propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and thelike. Examples of pharmaceutically acceptable, nontoxic acid additionsalts are salts of an amino group formed with inorganic acids such ashydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid andperchloric acid or with organic acids such as acetic acid, oxalic acid,maleic acid, tartaric acid, citric acid, succinic acid or malonic acidor by using other methods used in the art such as ion exchange Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate,butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. In some embodiments, organic acids from which salts can bederived include, for example, acetic acid, propionic acid, glycolicacid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinicacid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamicacid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like.

Pharmaceutically acceptable salts derived from appropriate bases includealkali metal, alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)⁴-salts.Inorganic bases from which salts can be derived include, but are notlimited to, sodium, potassium, lithium, ammonium, calcium, magnesium,iron, zinc, copper, manganese, aluminum, and the like. Organic basesfrom which salts can be derived include, but are not limited to,primary, secondary, and tertiary amines, substituted amines, includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like, examples include, but are not limited to,isopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, and ethanolamine. In some embodiments, thepharmaceutically acceptable base addition salt is ammonium, potassium,sodium, calcium, or magnesium salts. Representative alkali or alkalineearth metal salts include sodium, lithium, potassium, calcium,magnesium, iron, zinc, copper, manganese, aluminum, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, lower alkyl sulfonate and aryl sulfonate. Organic bases fromwhich salts can be derived include, for example, primary, secondary, andtertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines, basic ion exchange resins, and thelike, such as isopropylamine, trimethylamine, diethylamine,triethylamine, tripropylamine, and ethanolamine. In some embodiments,the pharmaceutically acceptable base addition salt is chosen fromammonium, potassium, sodium, calcium, and magnesium salts. Bis salts(i.e., two counterions) and higher salts (e.g., three or morecounterions) are encompassed within the meaning of pharmaceuticallyacceptable salts.

In addition, if a compound of the present disclosure is obtained as anacid addition salt, the free base can be obtained by basifying asolution of the acid salt. Conversely, if a product is a free base, anacid addition salt, particularly a pharmaceutically acceptable additionsalt, can be produced by dissolving the free base in a suitable organicsolvent and treating the solution with an acid, in accordance withconventional procedures for preparing acid addition salts from basecompounds. Those skilled in the art will recognize various syntheticmethodologies that can be used to prepare non-toxic pharmaceuticallyacceptable addition salts.

In certain embodiments, the pharmaceutically acceptable form is a“solvate” (e.g., a hydrate). As used herein, the term “solvate” refersto compounds that further include a stoichiometric or non-stoichiometricamount of solvent bound by non-covalent intermolecular forces. Thesolvate can be of a disclosed compound or a pharmaceutically acceptablesalt thereof. Where the solvent is water, the solvate is a “hydrate”.Pharmaceutically acceptable solvates and hydrates are complexes that,for example, can include 1 to about 100, or 1 to about 10, or one toabout 2, 3 or 4, solvent or water molecules. In some embodiments, thesolvate can be a channel solvate. It will be understood that the term“compound” as used herein encompasses the compound and solvates of thecompound, as well as mixtures thereof.

As used herein, and unless otherwise specified, “prodrug” is meant toindicate a compound that can be converted under physiological conditionsor by solvolysis to a biologically active compound described herein.Thus, the term “prodrug” refers to a precursor of a biologically activecompound that is pharmaceutically acceptable. A prodrug can be inactivewhen administered to a subject, but is converted in vivo to an activecompound, for example, by hydrolysis. In some embodiments, the prodrugcompound often offers advantages of solubility, tissue compatibility ordelayed release in a mammalian organism (see, e.g., Bundgard, H., Designof Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam). A discussionof prodrugs is provided in Higuchi, T., et al., “Pro-drugs as NovelDelivery Systems,” A.C.S. Symposium Series, Vol. 14, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated in full by reference herein. The term “prodrug” is alsomeant to include any covalently bonded carriers, which release theactive Formula (I) in vivo when such prodrug is administered to amammalian subject. Prodrugs of an active compound, as described herein,can be prepared by modifying functional groups present in the activeFormula (I) in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent active compound. Prodrugsinclude compounds wherein a hydroxy, amino or mercapto group is bondedto any group that, when the prodrug of the active Formula (I) isadministered to a mammalian subject, cleaves to form a free hydroxy,free amino or free mercapto group, respectively. Examples of prodrugsinclude, but are not limited to, acetate, formate, and benzoatederivatives of an alcohol; or acetamide, formamide, and benzamidederivatives of an amine functional group in the active compound, and thelike. Other examples of prodrugs include compounds that comprise —NO,—NO₂, —ONO, or —ONO₂ moieties. Prodrugs can typically be prepared usingwell-known methods, such as those described in Burger's MedicinalChemistry and Drug Discovery, 172-178, 949-982 (Manfred E. Wolff ed.,5th ed., 1995), and Design of Prodrugs (H. Bundgaard ed., Elselvier, NewYork, 1985).

For example, if a disclosed compound or a pharmaceutically acceptableform of the compound contains a carboxylic acid functional group, aprodrug can comprise a pharmaceutically acceptable ester formed by thereplacement of the hydrogen atom of the acid group with a group such as(C₁-C₈)alkyl, (C₂-C₁₂)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl havingfrom 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to 6 carbonatoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms,N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N—(C₁-C₂)alkylamino(C₂-C₃)alkyl (such as β-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N,N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl.

Similarly, if a disclosed compound or a pharmaceutically acceptable formof the compound contains an alcohol functional group, a prodrug can beformed by the replacement of the hydrogen atom of the alcohol group witha group such as (C₁-C₆)alkanoyloxymethyl, 1-((C₁-C₆)alkanoyloxy)ethyl,1-methyl-1-((C₁-C₆)alkanoyloxy)ethyl(C₁-C₆)alkoxycarbonyloxymethyl,N—(C₁-C₆)alkoxycarbonylaminomethyl, succinoyl, (C₁-C₆)alkanoyl,α-amino(C₁-C₄)alkanoyl, arylacyl and α-aminoacyl, orα-aminoacyl-α-aminoacyl, where each α-aminoacyl group is independentlyselected from the naturally occurring L-amino acids, P(O)(OH)₂,—P(O)(O(C₁-C₆)alkyl)₂ or glycosyl (the radical resulting from theremoval of a hydroxyl group of the hemiacetal form of a carbohydrate).

If a disclosed compound or a pharmaceutically acceptable form of theFormula (I) incorporates an amine functional group, a prodrug can beformed by the replacement of a hydrogen atom in the amine group with agroup such as R-carbonyl, RO-carbonyl, NRR′-carbonyl where R and R′ areeach independently (C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl, benzyl, a naturalα-aminoacyl or natural α-aminoacyl-natural α-aminoacyl, —C(OH)C(O)OY¹wherein Y¹ is H, (C₁-C₆)alkyl or benzyl, —C(OY²)Y³ wherein Y² is (C₁-C₄)alkyl and Y³ is (C₁-C₆)alkyl, carboxy(C₁-C₆)alkyl, amino(C₁-C₄)alkyl ormono-N—or di-N,N—(C₁-C₆)alkylaminoalkyl, —C(Y⁴)Y⁵ wherein Y⁴ is H ormethyl and Y⁵ is mono-N—or di-N,N—(C₁-C₆)alkylamino, morpholino,piperidin-1-yl or pyrrolidin-1-yl.

In certain embodiments, the pharmaceutically acceptable form is anisomer. “Isomers” are different compounds that have the same molecularformula. “Stereoisomers” are isomers that differ only in the way theatoms are arranged in space. As used herein, the term “isomer” includesany and all geometric isomers and stereoisomers. For example, “isomers”include geometric double bond cis- and trans-isomers, also termed E- andZ-isomers; R- and S-enantiomers; diastereomers, (d)-isomers and(l)-isomers, racemic mixtures thereof; and other mixtures thereof, asfalling within the scope of this disclosure.

Substituents around a carbon-carbon double bond alternatively can bereferred to as “cis” or “trans,” where “cis” represents substituents onthe same side of the double bond and “trans” represents substituents onopposite sides of the double bond. The arrangement of substituentsaround a carbocyclic ring can also be designated as “cis” or “trans.”The term “cis” represents substituents on the same side of the plane ofthe ring, and the term “trans” represents substituents on opposite sidesof the plane of the ring. Mixtures of compounds wherein the substituentsare disposed on both the same and opposite sides of plane of the ringare designated “cis/trans.”

“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A mixture of a pair of enantiomers in anyproportion can be known as a “racemic” mixture. The term “(±)” is usedto designate a racemic mixture where appropriate. “Diastereoisomers” arestereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. The absolute stereochemistry is specifiedaccording to the Cahn-Ingold-Prelog R-S system. When a Formula (I) is anenantiomer, the stereochemistry at each chirogenic carbon can bespecified by either R or S. Resolved compounds whose absoluteconfiguration is unknown can be designated (+) or (−) depending on thedirection (dextro- or levorotatory) which they rotate plane polarizedlight at the wavelength of the sodium D line. Certain of the compoundsdescribed herein contain one or more asymmetric centers and can thusgive rise to enantiomers, diastereomers, and other stereoisomeric formsthat can be defined, in terms of absolute stereochemistry at eachasymmetric atom, as (R)— or (S)—. The present chemical entities,pharmaceutical compositions and methods are meant to include all suchpossible isomers, including racemic mixtures, optically substantiallypure forms and intermediate mixtures. Optically active (R)— and (S)—isomers can be prepared, for example, using chiral synthons or chiralreagents, or resolved using conventional techniques.

As used herein, and unless otherwise specified, the term“stereomerically pure” means a composition or substance that comprisesone stereoisomer of a compound and is substantially free of otherstereoisomers of that compound. For example, a stereomerically purecomposition of a compound having one chiral center will be substantiallyfree of the opposite enantiomer of the compound. A stereomerically purecomposition of a compound having two chiral centers will besubstantially free of other stereoisomers (e.g., diastereoisomers orenantiomers, or syn or anti isomers, or cis or trans isomers) of thecompound. A typical stereomerically pure compound comprises greater thanabout 80 percent by weight of one stereoisomer of the compound and lessthan about 20 percent by weight of other stereoisomers of the compound,greater than about 90 percent by weight of one stereoisomer of thecompound and less than about 10 percent by weight of the otherstereoisomers of the compound, greater than about 95 percent by weightof one stereoisomer of the compound and less than about 5 percent byweight of the other stereoisomers of the compound, or greater than about97 percent by weight of one stereoisomer of the compound and less thanabout 3 percent by weight of the other stereoisomers of the compound.

As used herein, and unless otherwise specified, the term“enantiomerically pure” means a stereomerically pure composition of acompound having one or more chiral center(s).

As used herein, and unless otherwise specified, the terms “enantiomericexcess” and “diastereomeric excess” are used interchangeably herein. Insome embodiments, compounds with a single stereocenter can be referredto as being present in “enantiomeric excess,” and those with at leasttwo stereocenters can be referred to as being present in “diastereomericexcess.” For example, the term “enantiomeric excess” is well known inthe art and is defined as:

${ee}_{a} = {\left( \frac{{{{conc}.\mspace{14mu}{of}}\mspace{14mu} a} - {{{conc}.\mspace{14mu}{of}}\mspace{14mu} b}}{{{{conc}.\mspace{14mu}{of}}\mspace{14mu} a} + {{{conc}.\mspace{14mu}{of}}\mspace{14mu} b}} \right) \times 100}$

Thus, the term “enantiomeric excess” is related to the term “opticalpurity” in that both are measures of the same phenomenon. The value ofee will be a number from 0 to 100, zero being racemic and 100 beingenantiomerically pure. A compound which in the past might have beencalled 98% optically pure is now more precisely characterized by 96% ee.A 90% ee reflects the presence of 95% of one enantiomer and 5% of theother(s) in the material in question.

Some compositions described herein contain an enantiomeric excess of atleast about 50%, 75%, 90%, 95%, or 99% of the S enantiomer. In otherwords, the compositions contain an enantiomeric excess of the Senantiomer over the R enantiomer. In other embodiments, somecompositions described herein contain an enantiomeric excess of at leastabout 50%, 75%, 90%, 95%, or 99% of the R enantiomer. In other words,the compositions contain an enantiomeric excess of the R enantiomer overthe S enantiomer.

For instance, an isomer/enantiomer can, in some embodiments, be providedsubstantially free of the corresponding enantiomer, and can also bereferred to as “optically enriched,” “enantiomerically enriched,”“enantiomerically pure” and “non-racemic,” as used interchangeablyherein. These terms refer to compositions in which the percent by weightof one enantiomer is greater than the amount of that one enantiomer in acontrol mixture of the racemic composition (e.g., greater than about 1:1by weight). For example, an enantiomerically enriched preparation of theS enantiomer, means a preparation of the compound having greater thanabout 50% by weight of the S enantiomer relative to the R enantiomer,such as at least about 75% by weight, further such as at least about 80%by weight. In some embodiments, the enrichment can be much greater thanabout 80% by weight, providing a “substantially enantiomericallyenriched,” “substantially enantiomerically pure” or a “substantiallynon-racemic” preparation, which refers to preparations of compositionswhich have at least about 85% by weight of one enantiomer relative toother enantiomer, such as at least about 90% by weight, and further suchas at least 95% by weight. In certain embodiments, the compound providedherein is made up of at least about 90% by weight of one enantiomer. Inother embodiments, the Formula (I) is made up of at least about 95%,98%, or 99% by weight of one enantiomer.

Enantiomers can be isolated from racemic mixtures by any method known tothose skilled in the art, including chiral high pressure liquidchromatography (HPLC), the formation and crystallization of chiralsalts, or prepared by asymmetric syntheses. See, for example,Enantiomers, Racemates and Resolutions (Jacques, Ed., WileyInterscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977);Stereochemistry of Carbon Compounds (E. L. Eliel, Ed., McGraw-Hill, N Y,1962); and Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972).

In certain embodiments, the pharmaceutically acceptable form is atautomer. As used herein, the term “tautomer” is a type of isomer thatincludes two or more interconvertable compounds resulting from at leastone formal migration of a hydrogen atom and at least one change invalency (e.g., a single bond to a double bond, a triple bond to a singlebond, or vice versa). “Tautomerization” includes prototropic orproton-shift tautomerization, which is considered a subset of acid-basechemistry. “Prototropic tautomerization” or “proton-shifttautomerization” involves the migration of a proton accompanied bychanges in bond order. The exact ratio of the tautomers depends onseveral factors, including temperature, solvent, and pH. Wheretautomerization is possible (e.g., in solution), a chemical equilibriumof tautomers can be reached. Tautomerizations (i.e., the reactionproviding a tautomeric pair) can be catalyzed by acid or base, or canoccur without the action or presence of an external agent. Exemplarytautomerizations include, but are not limited to, keto-to-enol;amide-to-imide; lactam-to-lactim; enamine-to-imine; and enamine-to-(adifferent) enamine tautomerizations. An example of keto-enoltautomerization is the interconversion of pentane-2,4-dione and4-hydroxypent-3-en-2-one tautomers. Another example of tautomerizationis phenol-keto tautomerization. Another example of phenol-ketotautomerization is the interconversion of pyridin-4-ol andpyridin-4(1H)-one tautomers.

As used herein, and unless otherwise specified, structures depictedherein are also meant to include compounds which differ only in thepresence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement of ahydrogen by a deuterium or tritium, or the replacement of a carbon by¹³C—or ¹⁴C-enriched carbon, or the replacement of a nitrogen by ¹³N- or¹⁵N-enriched nitrogen, or the replacement of an oxygen by ¹⁴O—, ¹⁵O—,¹⁷O—, or ¹⁸O-enriched oxygen, or the replacement of a chlorine by ³⁵Cl—,³⁶Cl—, or ³⁷Cl-enriched chlorine, are within the scope of thisdisclosure.

In one embodiment, the compounds of the present disclosure can alsocontain unnatural proportions of atomic isotopes at one or more of atomsthat constitute such compounds. For example, the compounds can beradiolabeled with radioactive isotopes, such as, for example, tritium(³H), iodine-125 (¹²⁵I), or carbon-14 (¹⁴C). Certainisotopically-labeled disclosed compounds (e.g., those labeled with ³Hand ¹⁴C) are useful in compound and/or substrate tissue distributionassays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C) isotopes canallow for ease of preparation and detectability. Further, substitutionwith heavier isotopes such as deuterium (i.e., ²H) can afford certaintherapeutic advantages resulting from greater metabolic stability (e.g.,increased in vivo half-life or reduced dosage requirements).Isotopically labeled disclosed compounds can generally be prepared bysubstituting an isotopically labeled reagent for a non-isotopicallylabeled reagent. In some embodiments, provided herein are compounds thatcan also contain unnatural proportions of atomic isotopes at one or moreof atoms that constitute such compounds. All isotopic variations ofcompounds of the present disclosure, whether radioactive or not, areencompassed within the scope of the present disclosure.

As used herein, and unless otherwise specified, the terms “solvent,”“organic solvent,” or “inert solvent” each mean a solvent inert underthe conditions of the reaction being described in conjunction therewith,including, without limitation, benzene, toluene, acetonitrile (“MeCN”),ethyl acetate (“EtOAc”), isopropyl acetate (“IPAc”), hexanes, heptanes,dioxane, tetrahydrofuran (“THF”), dimethylformamide (“DMF”),dimethylacetamide (“DMA”), chloroform, methylene chloride (“DCM”),diethyl ether, methanol (“MeOH”), butanol (“1-BuOH”), methyl t-butylether (“MTBE”, or “TBME”), 2-butanone (“MEK”), N-methylpyrrolidone(“NMP”), pyridine, and the like. Unless specified to the contrary, thesolvents used in reactions described herein are inert organic solvents.Unless specified to the contrary, for each gram of a limiting reagent,one cc (or mL) of solvent constitutes a volume equivalent.

As used herein, and unless otherwise specified, “pharmaceuticallyacceptable carrier” or “pharmaceutically acceptable excipient” includesany and all solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic and absorption delaying agents and the like.The use of such media and agents for pharmaceutically active substancesis known in the art. Except insofar as any conventional media or agentis incompatible with the active ingredient, its use in the therapeuticcompositions of the present disclosure is contemplated. Supplementaryactive ingredients can also be incorporated into the compositions.

As used herein and unless otherwise specified, the term “solid form” andrelated terms refer to a physical form which is not predominantly in aliquid or a gaseous state. Solid forms may be crystalline, amorphous ormixtures thereof. In particular embodiments, solid forms may be liquidcrystals.

In some embodiments, a solid form provided herein is a single componentor multiple component solid form. A “single-component” solid formcomprising a compound of a formula consists essentially of the compoundof the formula. A “multiple-component” solid form comprising a compoundof a formula comprises a significant quantity of one or more additionalspecies, such as ions and/or molecules, within the solid form. Forexample, a crystalline multiple-component solid form comprising acompound of a formula further comprises one or more speciesnon-covalently bonded at regular positions in the crystal lattice. Amultiple component solid form provided herein may be a co-crystal.

As used herein and unless otherwise specified, the term “crystalline”and related terms, when used to describe a substance, modification,material, component or product mean that the substance, modification,material, component or product is substantially crystalline asdetermined by X-ray diffraction. See, e.g., Remington: The Science andPractice of Pharmacy, 21^(st) edition, Lippincott, Williams and Wilkins,Baltimore, Md. (2005); The United States Pharmacopeia, 23^(rd) edition,1843-1844 (1995).

As used herein and unless otherwise specified, the term “crystal forms”and related terms refer to solid forms that are crystalline. Crystalforms include single-component crystal forms and multiple-componentcrystal forms, and include, but are not limited to, polymorphs,solvates, hydrates, and other molecular complexes, as well as salts,solvates of salts, hydrates of salts, other molecular complexes ofsalts, and polymorphs thereof. In certain embodiments, a crystal form ofa substance may be substantially free of amorphous forms and/or othercrystal forms. In certain embodiments, a crystal form of a substance maycontain less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%,20%, 25%, 30%, 35%, 40%, 45% or 50% of one or more amorphous formsand/or other crystal forms on a weight basis. In certain embodiments, acrystal form of a substance may be physically and/or chemically pure. Incertain embodiments, a crystal form of a substance may be about 99%,98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/orchemically pure.

As used herein and unless otherwise specified, the terms “polymorphs,”“polymorphic forms” and related terms herein, refer to two or morecrystal forms that consist essentially of the same molecule, moleculesor ions. Like different crystal forms, different polymorphs may havedifferent physical properties such as, for example, meltingtemperatures, heats of fusion, solubilities, dissolution rates and/orvibrational spectra, as a result of the arrangement or conformation ofthe molecules and/or ions in the crystal lattice. The differences inphysical properties may affect pharmaceutical parameters such as storagestability, compressibility and density (important in formulation andproduct manufacturing), and dissolution rate (an important factor inbioavailability). Differences in stability can result from changes inchemical reactivity (e.g., differential oxidation, such that a dosageform discolors more rapidly when comprised of one polymorph than whencomprised of another polymorph) or mechanical changes (e.g., tabletscrumble on storage as a kinetically favored polymorph converts to athermodynamically more stable polymorph) or both (e.g., tablets of onepolymorph are more susceptible to breakdown at high humidity). As aresult of solubility/dissolution differences, in the extreme case, somesolid-state transitions may result in lack of potency or, at the otherextreme, toxicity. In addition, the physical properties may be importantin processing (for example, one polymorph might be more likely to formsolvates or might be difficult to filter and wash free of impurities,and particle shape and size distribution might be different betweenpolymorphs).

As used herein and unless otherwise specified, the term “solvate” and“solvated,” refer to a crystal form of a substance which containssolvent. The term “hydrate” and “hydrated” refer to a solvate whereinthe solvent comprises water. “Polymorphs of solvates” refers to theexistence of more than one crystal form for a particular solvatecomposition. Similarly, “polymorphs of hydrates” refers to the existenceof more than one crystal form for a particular hydrate composition. Theterm “desolvated solvate,” as used herein, refers to a crystal form of asubstance which may be prepared by removing the solvent from a solvate.

As used herein and unless otherwise specified, the term “amorphous,”“amorphous form,” and related terms used herein, mean that thesubstance, component or product in question is not substantiallycrystalline as determined by X-ray diffraction. In particular, the term“amorphous form” describes a disordered solid form, i.e., a solid formlacking long range crystalline order. In certain embodiments, anamorphous form of a substance may be substantially free of otheramorphous forms and/or crystal forms. In other embodiments, an amorphousform of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of one or more other amorphousforms and/or crystal forms on a weight basis. In certain embodiments, anamorphous form of a substance may be physically and/or chemically pure.In certain embodiments, an amorphous form of a substance may be about99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/orchemically pure.

Techniques for characterizing solid forms include, but are not limitedto, thermal gravimetric analysis (TGA), differential scanningcalorimetry (DSC), X-ray powder diffractometry (XRPD), gravimetric vaporsorption (GVS), single-crystal X-ray diffractometry, vibrationalspectroscopy, e.g., infrared (IR) and Raman spectroscopy, solid-stateand solution nuclear magnetic resonance (NMR) spectroscopy, opticalmicroscopy, hot stage optical microscopy, scanning electron microscopy(SEM), electron crystallography and quantitative analysis, particle sizeanalysis (PSA), surface area analysis, solubility measurements,dissolution measurements, elemental analysis and Karl Fischer analysis.Characteristic unit cell parameters may be determined using one or moretechniques such as, but not limited to, X-ray diffraction and neutrondiffraction, including single-crystal diffraction and powderdiffraction. Techniques useful for analyzing powder diffraction datainclude profile refinement, such as Rietveld refinement, which may beused, e.g., to analyze diffraction peaks associated with a single phasein a sample comprising more than one solid phase. Other methods usefulfor analyzing powder diffraction data include unit cell indexing, whichallows one of skill in the art to determine unit cell parameters from asample comprising crystalline powder.

In some embodiments, the solid forms, e.g., crystal forms, describedherein are substantially pure, i.e., substantially free of other solidforms and/or of other chemical compounds, containing less than about25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%,0.25% or 0.1% percent by weight of one or more other solid forms and/orof other chemical compounds.

Solid forms may exhibit distinct physical characterization data that areunique to a particular solid form, such as the crystal forms describedherein. These characterization data may be obtained by varioustechniques known to those skilled in the art, including for exampleX-ray powder diffraction, differential scanning calorimetry, thermalgravimetric analysis, and nuclear magnetic resonance spectroscopy. Thedata provided by these techniques may be used to identify a particularsolid form. One skilled in the art can determine whether a solid form isone of the forms described herein by performing one of thesecharacterization techniques and determining whether the resulting data“matches” the reference data provided herein, which is identified asbeing characteristic of a particular solid form. Characterization datathat “matches” those of a reference solid form is understood by thoseskilled in the art to correspond to the same solid form as the referencesolid form. In analyzing whether data “match,” a person of ordinaryskill in the art understands that particular characterization datapoints may vary to a reasonable extent while still describing a givensolid form, due to, for example, experimental error and routinesample-to-sample analysis.

The solid forms provided herein may be crystalline or an intermediateform (e.g., a mixture of crystalline and amorphous forms). The crystalforms described herein, therefore, may have varying degrees ofcrystallinity or lattice order. The solid forms described herein are notlimited by any particular degree of crystallinity or lattice order, andmay be 0-100% crystalline. Methods of determining the degree ofcrystallinity are known to those of ordinary skill in the art, such asthose described in Suryanarayanan, R., X-Ray Power Diffractometry,Physical Characterization of Pharmaceutical Salts, H. G. Brittain,Editor, Mercel Dekkter, Murray Hill, N.J., 1995, pp. 187-199, which isincorporated herein by reference in its entirety. In some embodiments,the solid forms described herein are about 0, 5, 10, 15, 20, 25, 30, 35,40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100% crystalline.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. For purposes of this application, thechemical elements are identified in accordance with the Periodic Tableof the Elements, CAS version, Handbook of Chemistry and Physics, 75^(th)Ed., inside cover, and specific functional groups are generally definedas described therein. Additionally, general principles of organicchemistry, as well as specific functional moieties and reactivity, aredescribed in Organic Chemistry, Thomas Sorrell, University ScienceBooks, Sausalito, 1999; Smith and March March's Advanced OrganicChemistry, 5^(th) Edition, John Wiley & Sons, Inc., New York, 2001;Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., NewYork, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3^(rd)Edition, Cambridge University Press, Cambridge, 1987.

The term “alkyl,” as used herein, refers to saturated, straight- orbranched-chain optionally substituted hydrocarbon radicals derived froman aliphatic moiety containing between one and six carbon atoms (e.g.,C₁₋₆ alkyl) by removal of a single hydrogen atom. In some embodiments,the alkyl group employed contains 1-5 carbon atoms. In anotherembodiment, the alkyl group employed contains 1-4 carbon atoms. In stillother embodiments, the alkyl group contains 1-3 carbon atoms. In yetanother embodiments, the alkyl group contains 1-2 carbons. Examples ofalkyl radicals include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl, iso-pentyl,tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl,n-decyl, n-undecyl, dodecyl, and the like.

The term “alkenyl,” as used herein, denotes a monovalent group derivedfrom a straight- or branched-chain optionally substituted aliphaticmoiety having at least one carbon-carbon double bond by the removal of asingle hydrogen atom. In certain embodiments, the alkenyl group contains2-6 carbon atoms (e.g., C₂₋₆ alkenyl). In certain embodiments, thealkenyl group contains 2-5 carbon atoms. In some embodiments, thealkenyl group contains 2-4 carbon atoms. In another embodiment, thealkenyl group employed contains 2-3 carbon atoms. Alkenyl groupsinclude, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,and the like.

The term “alkynyl,” as used herein, refers to a monovalent group derivedfrom a straight- or branched-chain optionally substituted aliphaticmoiety having at least one carbon-carbon triple bond by the removal of asingle hydrogen atom. In certain embodiments, the alkynyl group contains2-6 carbon atoms (e.g., C₂₋₆ alkynyl). In certain embodiments, thealkynyl group contains 2-5 carbon atoms. In some embodiments, thealkynyl group contains 2-4 carbon atoms. In another embodiment, thealkynyl group contains 2-3 carbon atoms. Representative alkynyl groupsinclude, but are not limited to, ethynyl, 2-propynyl (propargyl),1-propynyl, and the like.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic andbicyclic optionally substituted ring systems having a total of five totwelve ring members, wherein at least one ring in the system is aromaticand wherein each ring in the system contains three to seven ringmembers. In some embodiments, “aryl” refers to monocyclic and bicyclicoptionally substituted ring systems having a total of six to twelve ringmembers (e.g., C₆₋₁₂ aryl), wherein at least one ring in the system isaromatic and wherein each ring in the system contains three to sevenring members. The term “aryl” may be used interchangeably with the term“aryl ring”. In certain embodiments, “aryl” refers to an aromatic ringsystem which includes, but not limited to, phenyl, biphenyl, naphthyl,anthracyl and the like, which may bear one or more substituents. Alsoincluded within the scope of the term “aryl”, as it is used herein, is agroup in which an aromatic ring is fused to one or more non-aromaticrings, such as indanyl, phthalimidyl, naphthimidyl, phenantriidinyl, ortetrahydronaphthyl, and the like.

The terms “heteroaryl” used alone or as part of a larger moiety, e.g.,“heteroaralkyl”, or “heteroaralkoxy”, refer to optionally substitutedgroups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. In someembodiments, the term “heteroaryl” refers to optionally substitutedgroups as defined above having 6 to 10 ring atoms (e.g., C₆₋₁₂heteroaryl). The term “heteroatom” refers to nitrogen, oxygen, orsulfur, and includes any oxidized form of nitrogen or sulfur, and anyquaternized form of a basic nitrogen. Heteroaryl groups include, withoutlimitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl,isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl,pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. Theterms “heteroaryl” and “heteroar-”, as used herein, also include groupsin which a heteroaromatic ring is fused to one or more aryl,cycloaliphatic, or heterocyclyl rings, where the radical or point ofattachment is on the heteroaromatic ring. Non-limiting examples includeindolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl,indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl,cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl,carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, andpyrido[2,3-b]-1,4-oxazin-3(4H)-one. A heteroaryl group may be mono- orbicyclic. The term “heteroaryl” may be used interchangeably with theterms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any ofwhich terms include rings that are optionally substituted. The term“heteroaralkyl” refers to an alkyl group substituted by a heteroaryl,wherein the alkyl and heteroaryl portions independently are optionallysubstituted.

As described herein, compounds provided herein may contain “optionallysubstituted” moieties. In general, the term “substituted”, whetherpreceded by the term “optionally” or not, means that one or morehydrogens of the designated moiety are replaced with a suitablesubstituent. Unless otherwise indicated, an “optionally substituted”group may have a suitable substituent at each substitutable position ofthe group, and when more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this applicationare preferably those that result in the formation of stable orchemically feasible compounds. The term “stable”, as used herein, refersto compounds that are not substantially altered when subjected toconditions to allow for their production, detection, and, in certainembodiments, their recovery, purification, and use for one or more ofthe purposes disclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group are independently halogen;—(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘); —NO₂; —CN;—N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘); —N(R^(∘))C(S)R^(∘);—(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘) ₂;—(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃; —(CH₂)₀₋₄OC(O)R^(∘);—OC(O)(CH₂)₀₋₄SR—, SC(S)SR^(∘); —(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR^(∘)₂; —C(S)NR^(∘) ₂; —C(S)SR^(∘); —SC(S)SR^(∘), —(CH₂)₀₋₄OC(O)NR^(∘) ₂;—C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘);—C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘); —(CH₂)₀₋₄S(O)₂R^(∘);—(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘); —S(O)₂NR^(∘) ₂;—(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂; —N(R^(∘))S(O)₂R^(∘);—N(OR^(∘))R^(∘); —C(NH)NR^(∘) ₂; —P(O)₂R^(∘); —P(O)R^(∘) ₂; —OP(O)R^(∘)₂; —OP(O)(OR^(∘))₂; SiR^(∘) ₃; —(C₁₋₄ straight or branchedalkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substituted asdefined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur, or, notwithstanding the definition above, twoindependent occurrences of R^(∘), taken together with their interveningatom(s), form a 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur, which may be substituted as definedbelow.

Suitable monovalent substituents on R^(∘) or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), are independently halogen, —(CH₂)₀₋₂R^(●), -(haloR^(●)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), —(CH₂)₀₋₂CH(OR^(●))₂; —O(haloR^(●)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(●), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(●),—(CH₂)₀₋₂SR^(●), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(●),—(CH₂)₀₋₂NR^(●) ₂, —NO₂, —SiR^(●) ₃, —OSiR^(●) ₃, —C(O)SR^(●), —(C₁₋₄straight or branched alkylene)C(O)OR^(●), or —SSR^(●) wherein each R^(●)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O, ═S, ═NNR*₂,═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or—S(C(R*₂))₂₋₃S—, wherein each independent occurrence of R* is selectedfrom hydrogen, C₁₋₆ aliphatic which may be substituted as defined below,or an unsubstituted 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. Suitable divalent substituents that are bound tovicinal substitutable carbons of an “optionally substituted” groupinclude: —O(CR*₂)₂₋₃O—, wherein each independent occurrence of R* isselected from hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN, —C(O)OH,—C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein each R^(●) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein eachR^(†) is independently hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R^(†) are independentlyhalogen, —R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN,—C(O)OH, —C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein eachR^(●) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

5.2 Solid Forms

Potential pharmaceutical solids include crystalline solids and amorphoussolids. Amorphous solids are characterized by a lack of long-rangestructural order, whereas crystalline solids are characterized bystructural periodicity. The desired class of pharmaceutical soliddepends upon the specific application; amorphous solids are sometimesselected on the basis of, e.g., an enhanced dissolution profile, whilecrystalline solids may be desirable for properties such as, e.g.,physical or chemical stability (see, e.g., S. R. Vippagunta et al., Adv.Drug. Deliv. Rev., (2001) 48:3-26; L. Yu, Adv. Drug. Deliv. Rev., (2001)48:27-42). A change in solid form may affect a variety of physical andchemical properties, which may provide benefits or drawbacks inprocessing, formulation, stability and bioavailability, among otherimportant pharmaceutical characteristics.

Whether crystalline or amorphous, potential solid forms of apharmaceutical compound may include single-component andmultiple-component solids. Single-component solids consist essentiallyof the pharmaceutical compound in the absence of other compounds.Variety among single-component crystalline materials may potentiallyarise from the phenomenon of polymorphism, wherein multiplethree-dimensional arrangements exist for a particular pharmaceuticalcompound (see, e.g., S. R. Byrn et al., Solid State Chemistry of Drugs,(1999) SSCI, West Lafayette).

Additional diversity among the potential solid forms of a pharmaceuticalcompound may arise from the possibility of multiple-component solids.Crystalline solids comprising two or more ionic species are termed salts(see, e.g., Handbook of Pharmaceutical Salts: Properties, Selection andUse, P. H. Stahl and C. G. Wermuth, Eds., (2002), Wiley, Weinheim).Additional types of multiple-component solids that may potentially offerother property improvements for a pharmaceutical compound or saltthereof include, e.g., hydrates, solvates, co-crystals and clathrates,among others (see, e.g., S. R. Byrn et al., Solid State Chemistry ofDrugs, (1999) SSCI, West Lafayette). Provided herein are also cocrystalsof Compound 1 and polymorphs thereof. Multiple-component crystal formsmay potentially be susceptible to polymorphism, wherein a givenmultiple-component composition may exist in more than onethree-dimensional crystalline arrangement. The discovery of solid formsis of great importance in the development of a safe, effective, stableand marketable pharmaceutical compound.

The solid forms provided herein are useful as active pharmaceuticalingredients for the preparation of formulations for use in animals orhumans. Thus, embodiments herein encompass the use of these solid formsas a final drug product. Certain embodiments provide solid forms usefulin making final dosage forms with improved properties, e.g., powder flowproperties, compaction properties, tableting properties, stabilityproperties, and excipient compatibility properties, among others, thatare needed for manufacturing, processing, formulation and/or storage offinal drug products. Certain embodiments herein provide pharmaceuticalcompositions comprising a single-component crystal form, and/or amultiple-component crystal form comprising the compound of formula (I)and a pharmaceutically acceptable diluent, excipient or carrier.

Solid form and related terms refer to a physical form which is notpredominantly in a liquid or a gaseous state. Solid forms may becrystalline or mixtures of crystalline and amorphous forms. A“single-component”solid form comprising a particular compound consistsessentially of that compound. A “multiple-component” solid formcomprising a particular compound comprises that compound and asignificant quantity of one or more additional species, such as ionsand/or molecules, within the solid form. The solid forms provided hereinmay be crystalline or an intermediate form (e.g., a mixture ofcrystalline and amorphous forms). The crystal forms described herein,therefore, may have varying degrees of crystallinity or lattice order.The solid forms described herein are not limited to any particulardegree of crystallinity or lattice order, and may be 0-100% crystalline.Methods of determining the degree of crystallinity are known to those ofordinary skill in the art, such as those described in Suryanarayanan,R., X-Ray Powder Diffractometry, Physical Characterization ofPharmaceutical Solids, H. G. Brittain, Editor, Marcel Dekker, MurrayHill, N.J., 1995, pp. 187-199, which is incorporated herein by referencein its entirety. In some embodiments, the solid forms described hereinare about 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,80, 85, 90, 95 or 100% crystalline.

Solid forms may exhibit distinct physical characterization data that areunique to a particular solid form, such as the crystal forms describedherein. These characterization data may be obtained by varioustechniques known to those skilled in the art, including for exampleX-ray powder diffraction, differential scanning calorimetry, thermalgravimetric analysis, and nuclear magnetic resonance spectroscopy. Thedata provided by these techniques may be used to identify a particularsolid form. One skilled in the art can determine whether a solid form isone of the forms described herein by performing one of thesecharacterization techniques and determining whether the resulting datais “substantially similar” to the reference data provided herein, whichis identified as being characteristic of a particular solid form.Characterization data that is “substantially similar” to those of areference solid form is understood by those skilled in the art tocorrespond to the same solid form as the reference solid form. Inanalyzing whether data is “substantially similar,” a person of ordinaryskill in the art understands that particular characterization datapoints may vary to a reasonable extent while still describing a givensolid form, due to, for example, experimental error and routinesample-to-sample analysis.

In some embodiments, provided herein are solid forms comprising acompound of formula (I):

or a salt, solvate (e.g., hydrate), or solvate of a salt thereof, or amixture thereof. In one embodiment, the solid form comprising a compoundof formula (I) can be a crystalline form, a partially crystalline form,or a mixture of crystalline form(s) and amorphous form(s). In oneembodiment, provided herein is a solid form comprising a crystallineform of a compound of formula (I), or a salt, solvate (e.g., hydrate),or solvate of a salt thereof, or a mixture thereof. In one embodiment,the solid form further comprises a coformer. In one embodiment, thesolid form comprising Compound 1 and a coformer is a cocrystal. Inanother embodiment, the solid form is an amorphous form. In oneembodiment, the solid form is substantially pure. The compound offormula (I) has a chemical name of(S)-2-amino-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide.The compound of formula (I) is described in US2015/011874, the entiretyof which is incorporated herein by reference.

In some embodiments, the Formula (I) is a racemic mixture of (S)- and(R)-isomers. In other embodiments, provided herein is a mixture ofcompounds wherein individual compounds of the mixture existpredominately in an (S)- or (R)-isomeric configuration. For example, thecompound mixture has an (S)-enantiomeric excess of greater than about55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%,about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about99.5%, or more. In other embodiments, the compound mixture has an(S)-enantiomeric excess of greater than about 55% to about 99.5%,greater than about 60% to about 99.5%, greater than about 65% to about99.5%, greater than about 70% to about 99.5%, greater than about 75% toabout 99.5%, greater than about 80% to about 99.5%, greater than about85% to about 99.5%, greater than about 90% to about 99.5%, greater thanabout 95% to about 99.5%, greater than about 96% to about 99.5%, greaterthan about 97% to about 99.5%, greater than about 98% to greater thanabout 99.5%, greater than about 99% to about 99.5%, or more.

In other embodiments, the compound mixture has an (R)-enantiomericpurity of greater than about 55%, about 60%, about 65%, about 70%, about75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%,about 98%, about 99%, about 99.5% or more. In some other embodiments,the compound mixture has an (R)-enantiomeric excess of greater thanabout 55% to about 99.5%, greater than about 60% to about 99.5%, greaterthan about 65% to about 99.5%, greater than about 70% to about 99.5%,greater than about 75% to about 99.5%, greater than about 80% to about99.5%, greater than about 85% to about 99.5%, greater than about 90% toabout 99.5%, greater than about 95% to about 99.5%, greater than about96% to about 99.5%, greater than about 97% to about 99.5%, greater thanabout 98% to greater than about 99.5%, greater than about 99% to about99.5% or more.

5.2.1. Solid Forms of Compound 1

Provided herein is a solid form comprising a compound of Formula (I):

or a salt, or solvate (e.g., hydrate), or solvate of a salt thereof, ora mixture thereof.

In one embodiment, provided herein a solid form comprising a free baseof Compound 1, or a solvate (e.g., hydrate) thereof. In one embodiment,provided herein is a solid form comprising an anhydrous free base ofCompound 1. In one embodiment, provided herein is a solid formcomprising a solvate of a free base of Compound 1. In one embodiment,provided herein is a solid form comprising a hydrate of a free base ofCompound 1.

It is contemplated that Compound 1, or a salt, or solvate (e.g.,hydrate), or solvate of a salt thereof, or a mixture thereof, can existin a variety of solid forms. Such solid forms include crystalline solids(e.g., polymorphs of anhydride Compound 1, polymorphs of hydrates ofCompound 1, and polymorphs of solvates of Compound 1), amorphous solids,or mixtures of crystalline and amorphous solids. In one embodiment, thesolid form is substantially crystalline. In one embodiment, the solidform is crystalline.

In some embodiments, the molar ratio of Compound 1 to the solvent/waterin the solid form ranges from about 10:1 to about 1:10. In someembodiments, the molar ratio of Compound 1 to the solvent/water in thesolid form ranges from about 5:1 to about 1:5. In some embodiments, themolar ratio of Compound 1 to the solvent/water in the solid form rangesfrom about 3:1 to about 1:3. In some embodiments, the molar ratio ofCompound 1 to the solvent/water in the solid form ranges from about 2:1to about 1:2. In one embodiment, the molar ratio is about 1:2 (i.e.,bis-solvate/hydrate). In another embodiment, the molar ratio is about1:1 (i.e., mono-solvate/hydrate). In yet another embodiment, the molarratio is about 2:1 (i.e., hemi-solvate/hydrate).

5.2.1.1 Form 1 of Compound 1

In some embodiments, provided herein is Form 1 of a compound of formula(I). In one embodiment, Form 1 of Compound 1 is a crystallinenon-solvated anhydrous free base of Compound 1. In some embodiments,Form 1 of Compound 1 is substantially free of amorphous Compound 1. Insome embodiments, Form 1 of Compound 1 is substantially free of othercrystalline forms (i.e., polymorphs) of Compound 1. In some embodiments,Form 1 of Compound 1 is substantially free of salts of Compound 1. Insome embodiments, Form 1 of Compound 1 is provided as substantially pureForm 1 of Compound 1. In some embodiments, one or more residual solvent(e.g., small amount of EtOH or ^(i)PrOH) may be present in Form 1 ofCompound 1, but the residual solvent does not form a solvate of Compound1.

A representative XRPD pattern of Form 1 of Compound 1 is provided inFIG. 1.

In one embodiment, Form 1 has an X-ray powder diffraction (XRPD) patterncomprising peaks at 16.8, 23.6, and 25.6 degrees 2θ, plus or minus 0.2.In one embodiment, Form 1 has an XRPD pattern further comprising atleast one peak selected from 14.6 and 21.2 degrees 2θ, plus or minus0.2. In one embodiment, Form 1 has an XRPD pattern comprising peaks at14.6, 16.8, 21.2, 23.6, and 25.6 degrees 2θ, in combination with atleast one peak selected from 11.3, 15.4, 16.2, 18.4, 20.5, 22.6, 24.3,26.6, 27.1, and 29.5 degrees 2θ, plus or minus 0.2.

In one embodiment, Form 1 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 11.3, 14.6, 15.4, 16.2, 16.8, 18.4, 20.5, 21.2, 22.6, 23.6,24.3, 25.6, 26.6, 27.1, and 29.5 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 1 has an XRPD pattern substantially as shown inFIG. 1.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 1 ofCompound 1 is provided in FIG. 2.

In one embodiment, Form 1 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at about 255° C. and/ora peak temperature at about 257° C. In one embodiment, Form 1 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 2. In another embodiment, Form 1 exhibitsan endothermic event, as characterized by DSC, with an onset temperatureat about 242° C. and/or a peak temperature at about 251° C. In yetanother embodiment, Form 1 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at from about 242° C. toabout 255° C.

In one embodiment, Form 1 exhibits a weight loss of about 0.6% uponheating from about 230° C. to about 310° C. In one embodiment, Form 1 ischaracterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 2. In another embodiment, Form 1 exhibits aweight loss of about 0.4% upon heating from about 25° C. to about 70°C., and a weight loss of about 1.1% upon heating from about 200° C. toabout 280° C.

A representative gravimetric vapor sorption (GVS) isotherm of Form 1 ispresented in FIG. 3. In one embodiment, Form 1 exhibits a weightincrease of about 0.5% when subjected to an increase in relativehumidity from about 0 to about 90% relative humidity. In one embodiment,Form 1 is characterized by a GVS thermogram substantially as shown inthe GVS thermogram presented in FIG. 3.

In one embodiment, Form 1 has approximately unit cell dimensions of:a=11.1 Å, b=12.8 Å, c=16.1 Å, α=90°, β=90°, and γ=90°. In oneembodiment, Form 1 has approximately unit cell dimensions of: a=11.14 Å,b=12.76 Å, c=16.13 Å, α=90°, β=90°, and γ=90°. In one embodiment, Form 1has approximately unit cell dimensions of: a=11.140 Å, b=12.758 Å,c=16.131 Å, α=90°, β=90°, and γ=90°. In one embodiment, Form 1 has aunit cell of a space group of P2₁2₁2₁. In one embodiment, Form 1 has avolume of about 2292.5 Å³/cell. In one embodiment, Form 1 has a Z valueof 4. In one embodiment, Form 1 has a density of about 1.279 g/cm³.

In one embodiment, Form 1 is anhydrous. In one embodiment, Form 1 isnon-hygroscopic. In one embodiment, Form 1 is stable after storage at40° C./75% RH or 25° C./96% RH for more than 9 months.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.2 Form 2 of Compound 1

In some embodiments, provided herein is Form 2 of a compound of formula(I). In one embodiment, Form 2 of Compound 1 is a crystalline solvate offree base of Compound 1. In some embodiments, Form 2 of Compound 1 issubstantially free of amorphous Compound 1. In some embodiments, Form 2of Compound 1 is substantially free of other crystalline forms (i.e.,polymorphs) of Compound 1. In some embodiments, Form 2 of Compound 1 issubstantially free of salts of Compound 1. In some embodiments, Form 2of Compound 1 is provided as substantially pure Form 2 of Compound 1.

In one embodiment, the molar ratio of Compound 1 to the solvent in Form2 ranges from about 1:0.5 to about 1:2. In one embodiment, the molarratio of Compound 1 to the solvent in Form 2 ranges from about 1:0.75 toabout 1:1.25. In one embodiment, the molar ratio of Compound 1 to thesolvent in Form 2 ranges from about 1:0.75 to about 1:1. In oneembodiment, the molar ratio of Compound 1 to the solvent in Form 2 isabout 1:0.85. In one embodiment, the molar ratio of Compound 1 to thesolvent in Form 2 is about 1:1.

In one embodiment, Form 2 is an acetone/DCM solvate of free base ofCompound 1. In one embodiment, the molar ratio of Compound 1:acetone:DCMin Form 2 is about 1:0.1:0.75. In another embodiment, Form 2 is an1-propanol solvate of free base of Compound 1. In one embodiment, themolar ratio of Compound 1 to 1-propanol in Form 2 is about 1:0.85. Inanother embodiment, Form 2 is an DCM solvate of free base of Compound 1.In one embodiment, the molar ratio of Compound 1 to DCM in Form 2 isabout 1:1.

A representative XRPD pattern of Form 2 of Compound 1 is provided inFIG. 4. Another representative XRPD pattern of Form 2 of Compound 1 isprovided in FIG. 8.

In one embodiment, Form 2 has an XRPD pattern comprising peaks at 13.6,14.9, and 21.0 degrees 2θ, plus or minus 0.2. In one embodiment, Form 2has an XRPD pattern further comprising at least one peak selected from7.4 and 16.7 degrees 2θ, plus or minus 0.2. In one embodiment, Form 2has an XRPD pattern comprising peaks at 7.4, 13.6, 14.9, 16.7, and 21.0degrees 2θ, in combination with at least one peak selected from 9.5,18.1, 18.4, 19.7, 20.8, 22.4, 23.2, 24.5, 26.2, and 26.8 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 2 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 7.4, 9.5, 13.6, 14.9, 16.7, 18.1, 18.4, 19.7, 20.8, 21.0,22.4, 23.2, 24.5, 26.2, and 26.8 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 2 has an XRPD pattern substantially as shown inFIG. 4. In another embodiment, Form 2 has an XRPD pattern substantiallyas shown in FIG. 8.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 2 ofCompound 1 is provided in FIG. 5.

In one embodiment, Form 2 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at about 168° C. and/ora peak temperature at about 182° C. In one embodiment, Form 2 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 5.

In one embodiment, Form 2 exhibits a weight loss of about 12.9% uponheating from about 80° C. to about 240° C. In one embodiment, Form 2 ischaracterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 5.

Another representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 2 ofCompound 1 is provided in FIG. 7.

In one embodiment, Form 2 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 25° C., anendothermic event with an onset temperature at about 151° C., anexothermic event with an onset temperature at about 179° C., or anendothermic event with an onset temperature at about 244° C. In oneembodiment, Form 2 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 25° C., an endothermic eventwith an onset temperature at about 151° C., an exothermic event with anonset temperature at about 179° C., and an endothermic event with anonset temperature at about 244° C. In one embodiment, Form 2 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 7.

In one embodiment, Form 2 exhibits a weight loss of about 0.9% uponheating from about 25° C. to about 75° C., and a weight loss of about8.5% upon heating from about 75° C. to about 250° C. In one embodiment,Form 2 is characterized by a TGA thermogram substantially as shown inthe TGA thermogram presented in FIG. 7.

A representative gravimetric vapor sorption (GVS) isotherm of Form 2 ispresented in FIG. 6. In one embodiment, Form 2 exhibits a weightincrease of about 1.3% when subjected to an increase in relativehumidity from about 0 to about 90% relative humidity. In one embodiment,Form 2 is characterized by a GVS thermogram substantially as shown inthe GVS thermogram presented in FIG. 6.

In one embodiment, Form 2 is non-hygroscopic.

In one embodiment, Form 2 has approximately unit cell dimensions of:a=8.7 Å, b=13.2 Å, c=26.0 Å, α=90°, β=90°, and γ=90°. In one embodiment,Form 2 has approximately unit cell dimensions of: a=8.73 Å, b=13.22 Å,c=25.96 Å, α=90°, β=90°, and γ=90°. In one embodiment, Form 2 hasapproximately unit cell dimensions of: a=8.729 Å, b=13.222 Å, c=25.955Å, α=90°, β=90°, and γ=90°. In one embodiment, Form 2 has a unit cell ofa space group of P2₁2₁2₁. In one embodiment, Form 2 has a volume ofabout 2995.6 Å³/cell. In one embodiment, Form 2 has a Z value of 4. Inone embodiment, Form 2 has a density of about 1.360 Mg/m³.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.3 Form 3 of Compound 1

In some embodiments, provided herein is Form 3 of a compound of formula(I). In one embodiment, Form 3 of Compound 1 is a crystalline solvate offree base of Compound 1. In some embodiments, Form 3 of Compound 1 issubstantially free of amorphous Compound 1. In some embodiments, Form 3of Compound 1 is substantially free of other crystalline forms (i.e.,polymorphs) of Compound 1. In some embodiments, Form 3 of Compound 1 issubstantially free of salts of Compound 1. In some embodiments, Form 3of Compound 1 is provided as substantially pure Form 3 of Compound 1.

In one embodiment, the molar ratio of Compound 1 to the solvent in Form3 ranges from about 1:0.2 to about 1:1. In one embodiment, Form 3 is a2-methyl-1-propanol solvate of free base of Compound 1. In oneembodiment, the molar ratio of Compound 1 to 2-methyl-1-propanol in Form3 is about 1:0.79. In another embodiment, Form 3 is a MEK solvate offree base of Compound 1. In one embodiment, the molar ratio of Compound1 to MEK in Form 3 is about 1:0.25.

A representative XRPD pattern of Form 3 of Compound 1 is provided inFIG. 9.

In one embodiment, Form 3 has an XRPD pattern comprising peaks at 17.9,20.6, and 25.8 degrees 2θ, plus or minus 0.2. In one embodiment, Form 3has an XRPD pattern further comprising at least one peak selected from11.7 and 23.5 degrees 2θ, plus or minus 0.2. In one embodiment, Form 3has an XRPD pattern comprising peaks at 11.7, 17.9, 20.6, 23.5, and 25.8degrees 2θ, in combination with at least one peak selected from 7.4,10.2, 13.5, 19.3, 19.5, 21.0, 21.5, 22.4, 23.7, and 26.5 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 3 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 7.4, 10.2, 11.7, 13.5, 17.9, 19.3, 19.5, 20.6, 21.0, 21.5,22.4, 23.5, 23.7, 25.8, and 26.5 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 3 has an XRPD pattern substantially as shown inFIG. 9.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 3 ofCompound 1 is provided in FIG. 10.

In one embodiment, Form 3 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 29° C., anendothermic event with an onset temperature at about 126° C., anendothermic event with an onset temperature at about 148° C., anexothermic event with an onset temperature at about 181° C., or anendothermic event with an onset temperature at about 246° C. In oneembodiment, Form 3 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 29° C., an endothermic eventwith an onset temperature at about 126° C., an endothermic event with anonset temperature at about 148° C., an exothermic event with an onsettemperature at about 181° C., and an endothermic event with an onsettemperature at about 246° C. In one embodiment, Form 3 is characterizedby a DSC thermogram substantially as shown in the DSC thermogrampresented in FIG. 10.

In one embodiment, Form 3 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 30° C., anendothermic event with an onset temperature at about 127° C., anendothermic event with an onset temperature at about 137° C., anexothermic event with an onset temperature at about 169° C., anendothermic event with an onset temperature at about 207° C., or anendothermic event with an onset temperature at about 250° C. In oneembodiment, Form 3 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 30° C., an endothermic eventwith an onset temperature at about 127° C., an endothermic event with anonset temperature at about 137° C., an exothermic event with an onsettemperature at about 169° C., an endothermic event with an onsettemperature at about 207° C., and an endothermic event with an onsettemperature at about 250° C.

In one embodiment, Form 3 exhibits a weight loss of about 0.8% uponheating from about 25° C. to about 75° C., and a weight loss of about10.4% upon heating from about 75° C. to about 300° C. In one embodiment,Form 3 is characterized by a TGA thermogram substantially as shown inthe TGA thermogram presented in FIG. 10.

In one embodiment, Form 3 exhibits a weight loss of about 2.0% uponheating from about 25° C. to about 80° C., and a weight loss of about3.4% upon heating from about 80° C. to about 175° C.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.4 Form 4 of Compound 1

In some embodiments, provided herein is Form 4 of a compound of formula(I). In one embodiment, Form 4 of Compound 1 is a crystalline solvate offree base of Compound 1. In some embodiments, Form 4 of Compound 1 issubstantially free of amorphous Compound 1. In some embodiments, Form 4of Compound 1 is substantially free of other crystalline forms (i.e.,polymorphs) of Compound 1. In some embodiments, Form 4 of Compound 1 issubstantially free of salts of Compound 1. In some embodiments, Form 4of Compound 1 is provided as substantially pure Form 4 of Compound 1.

In one embodiment, the molar ratio of Compound 1 to the solvent in Form4 ranges from about 1:0.75 to about 1:1. In one embodiment, the molarratio of Compound 1 to the solvent in Form 4 ranges from about 1:0.83 toabout 1:0.9. In one embodiment, Form 4 is an isopropyl alcohol solvateof free base of Compound 1. In one embodiment, the molar ratio ofCompound 1 to isopropyl alcohol in Form 4 is about 1:0.9. In anotherembodiment, the molar ratio of Compound 1 to isopropyl alcohol in Form 4is about 1:0.83.

A representative XRPD pattern of Form 4 of Compound 1 is provided inFIG. 11.

In one embodiment, Form 4 has an XRPD pattern comprising peaks at 7.4,18.0, and 20.7 degrees 2θ, plus or minus 0.2. In one embodiment, Form 4has an XRPD pattern further comprising at least one peak selected from11.9 and 13.6 degrees 2θ, plus or minus 0.2. In one embodiment, Form 4has an XRPD pattern comprising peaks at 7.4, 11.9, 13.6, 18.0, and 20.7degrees 2θ, in combination with at least one peak selected from 10.3,19.3, 19.6, 19.8, 21.0, 21.8, 23.6, 23.8, 26.0, and 26.6 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 4 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 7.4, 10.3, 11.9, 13.6, 18.0, 19.3, 19.6, 19.8, 20.7, 21.0,21.8, 23.6, 23.8, 26.0, and 26.6 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 4 has an XRPD pattern substantially as shown inFIG. 11.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 4 ofCompound 1 is provided in FIG. 12.

In one embodiment, Form 4 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 30° C., anendothermic event with an onset temperature at about 156° C., anexothermic event with an onset temperature at about 190° C., or anendothermic event with an onset temperature at about 245° C. In oneembodiment, Form 4 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 30° C., an endothermic eventwith an onset temperature at about 156° C., an exothermic event with anonset temperature at about 190° C., and an endothermic event with anonset temperature at about 245° C. In one embodiment, Form 4 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 12.

In one embodiment, Form 4 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 28° C., anendothermic event with an onset temperature at about 156° C., anexothermic event with an onset temperature at about 201° C., or anendothermic event with an onset temperature at about 247° C. In oneembodiment, Form 4 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 28° C., an endothermic eventwith an onset temperature at about 156° C., an exothermic event with anonset temperature at about 201° C., and an endothermic event with anonset temperature at about 247° C.

In one embodiment, Form 4 exhibits a weight loss of about 0.7% uponheating from about 25° C. to about 75° C., and a weight loss of about9.3% upon heating from about 75° C. to about 225° C. In one embodiment,Form 4 is characterized by a TGA thermogram substantially as shown inthe TGA thermogram presented in FIG. 12.

In one embodiment, Form 4 exhibits a weight loss of about 0.8% uponheating from about 25° C. to about 75° C., and a weight loss of about8.6% upon heating from about 75° C. to about 250° C.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.5 Form 5 of Compound 1

In some embodiments, provided herein is Form 5 of a compound of formula(I). In one embodiment, Form 5 of Compound 1 is a crystalline solvate offree base of Compound 1. In some embodiments, Form 5 of Compound 1 issubstantially free of amorphous Compound 1. In some embodiments, Form 5of Compound 1 is substantially free of other crystalline forms (i.e.,polymorphs) of Compound 1. In some embodiments, Form 5 of Compound 1 issubstantially free of salts of Compound 1. In some embodiments, Form 5of Compound 1 is provided as substantially pure Form 5 of Compound 1.

In one embodiment, the molar ratio of Compound 1 to the solvent in Form5 ranges from about 1:0.1 to about 1:0.2. In one embodiment, Form 5 isan anisole solvate of free base of Compound 1. In one embodiment, themolar ratio of Compound 1 to anisole in Form 5 is about 1:0.12.

A representative XRPD pattern of Form 5 of Compound 1 is provided inFIG. 13.

In one embodiment, Form 5 has an XRPD pattern comprising peaks at 21.0,22.1, and 25.2 degrees 2θ, plus or minus 0.2. In one embodiment, Form 5has an XRPD pattern further comprising at least one peak selected from14.5 and 19.2 degrees 2θ, plus or minus 0.2. In one embodiment, Form 5has an XRPD pattern comprising peaks at 14.5, 19.2, 21.0, 22.1, and 25.2degrees 2θ, in combination with at least one peak selected from 7.9,11.0, 12.7, 16.6, 18.0, 23.3, 27.7, 28.5, 29.1, and 29.2 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 5 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 7.9, 11.0, 12.7, 14.5, 16.6, 18.0, 19.2, 21.0, 22.1, 23.3,25.2, 27.7, 28.5, 29.1, and 29.2 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 5 has an XRPD pattern substantially as shown inFIG. 13.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 5 ofCompound 1 is provided in FIG. 14.

In one embodiment, Form 5 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 126° C., or anendothermic event with an onset temperature at about 254° C. In oneembodiment, Form 5 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 126° C., and an endothermicevent with an onset temperature at about 254° C. In one embodiment, Form5 is characterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 14.

In one embodiment, Form 5 exhibits a weight loss of about 2.1% uponheating from about 175° C. to about 300° C. In one embodiment, Form 5 ischaracterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 14.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.6 Form 6 of Compound 1

In some embodiments, provided herein is Form 6 of a compound of formula(I). In one embodiment, Form 6 of Compound 1 is a crystalline hydrate offree base of Compound 1. In some embodiments, Form 6 of Compound 1 issubstantially free of amorphous Compound 1. In some embodiments, Form 6of Compound 1 is substantially free of other crystalline forms (i.e.,polymorphs) of Compound 1. In some embodiments, Form 6 of Compound 1 issubstantially free of salts of Compound 1. In some embodiments, Form 6of Compound 1 is provided as substantially pure Form 6 of Compound 1.

In one embodiment, the molar ratio of Compound 1 to the water in Form 6ranges from about 1:2 to about 1:4. In one embodiment, the molar ratioof Compound 1 to the water in Form 6 is about 1:3.3.

A representative XRPD pattern of Form 6 of Compound 1 is provided inFIG. 15.

In one embodiment, Form 6 has an XRPD pattern comprising peaks at 4.8,19.9, and 26.7 degrees 2θ, plus or minus 0.2. In one embodiment, Form 6has an XRPD pattern further comprising at least one peak selected from11.9 and 24.8 degrees 2θ, plus or minus 0.2. In one embodiment, Form 6has an XRPD pattern comprising peaks at 4.8, 11.9, 19.9, 24.8, and 26.7degrees 2θ, in combination with at least one peak selected from 12.2,12.4, 14.1, 16.0, 17.7, 18.1, 18.9, 20.9, 24.0, and 27.1 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 6 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 4.8, 11.9, 12.2, 12.4, 14.1, 16.0, 17.7, 18.1, 18.9, 19.9,20.9, 24.0, 24.8, 26.7, and 27.1 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 6 has an XRPD pattern substantially as shown inFIG. 15.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 6 ofCompound 1 is provided in FIG. 16.

In one embodiment, Form 6 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 46° C., anendothermic event with an onset temperature at about 154° C., or anendothermic event with an onset temperature at about 243° C. In oneembodiment, Form 6 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 46° C., an endothermic eventwith an onset temperature at about 154° C., and an endothermic eventwith an onset temperature at about 243° C. In one embodiment, Form 6 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 16.

In one embodiment, Form 6 exhibits a weight loss of about 10.3% uponheating from about 30° C. to about 100° C. In one embodiment, Form 6 ischaracterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 16.

A representative gravimetric vapor sorption (GVS) isotherm of Form 6 ispresented in FIG. 17. In one embodiment, Form 6 exhibits a weightincrease of about 14% when subjected to an increase in relative humidityfrom about 0 to about 90% relative humidity. In one embodiment, Form 6is characterized by a GVS thermogram substantially as shown in the GVSthermogram presented in FIG. 17.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.7 Form 7 of Compound 1

In some embodiments, provided herein is Form 7 of a compound of formula(I). In some embodiments, Form 7 of Compound 1 is substantially free ofamorphous Compound 1. In some embodiments, Form 7 of Compound 1 issubstantially free of other crystalline forms (i.e., polymorphs) ofCompound 1. In some embodiments, Form 7 of Compound 1 is substantiallyfree of salts of Compound 1. In some embodiments, Form 7 of Compound 1is provided as substantially pure Form 7 of Compound 1.

A representative XRPD pattern of Form 7 of Compound 1 is provided inFIG. 18.

In one embodiment, Form 7 has an XRPD pattern comprising peaks at 7.5,12.3, and 20.7 degrees 2θ, plus or minus 0.2. In one embodiment, Form 7has an XRPD pattern further comprising at least one peak selected from13.7 and 17.2 degrees 2θ, plus or minus 0.2. In one embodiment, Form 7has an XRPD pattern comprising peaks at 7.5, 12.3, 13.7, 17.2, and 20.7degrees 2θ, in combination with at least one peak selected from 11.8,14.9, 18.0, 18.4, 19.6, 20.2, 21.1, 23.5, 23.6, and 25.9 degrees 2θ,plus or minus 0.2.

In one embodiment, Form 7 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 7.5, 11.8, 12.3, 13.7, 14.9, 17.2, 18.0, 18.4, 19.6, 20.2,20.7, 21.1, 23.5, 23.6, and 25.9 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 7 has an XRPD pattern substantially as shown inFIG. 18.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form 7 ofCompound 1 is provided in FIG. 19.

In one embodiment, Form 7 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 30° C., anendothermic event with an onset temperature at about 127° C., anendothermic event with an onset temperature at about 137° C., anexothermic event with an onset temperature at about 169° C., anendothermic event with an onset temperature at about 207° C., or anendothermic event with an onset temperature at about 250° C. In oneembodiment, Form 7 exhibits, as characterized by DSC, an endothermicevent with an onset temperature at about 30° C., an endothermic eventwith an onset temperature at about 127° C., an endothermic event with anonset temperature at about 137° C., an exothermic event with an onsettemperature at about 169° C., an endothermic event with an onsettemperature at about 207° C., and an endothermic event with an onsettemperature at about 250° C. In one embodiment, Form 7 is characterizedby a DSC thermogram substantially as shown in the DSC thermogrampresented in FIG. 19.

In one embodiment, Form 7 exhibits a weight loss of about 2.0% uponheating from about 10° C. to about 90° C., and a weight loss of about3.4% upon heating from about 90° C. to about 190° C. In one embodiment,Form 7 is characterized by a TGA thermogram substantially as shown inthe TGA thermogram presented in FIG. 19.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.1.8 Form 8 of Compound 1

In some embodiments, provided herein is Form 8 of a compound of formula(I). In some embodiments, Form 8 of Compound 1 is substantially free ofamorphous Compound 1. In some embodiments, Form 8 of Compound 1 issubstantially free of other crystalline forms (i.e., polymorphs) ofCompound 1. In some embodiments, Form 8 of Compound 1 is substantiallyfree of salts of Compound 1. In some embodiments, Form 8 of Compound 1is provided as substantially pure Form 8 of Compound 1.

A representative XRPD pattern of Form 8 of Compound 1 is provided inFIG. 20.

In one embodiment, Form 8 has an XRPD pattern comprising peaks at 18.8,20.8, and 24.5 degrees 2θ, plus or minus 0.2. In one embodiment, Form 8has an XRPD pattern further comprising at least one peak selected from16.0 and 17.9 degrees 2θ, plus or minus 0.2. In one embodiment, Form 8has an XRPD pattern comprising peaks at 16.0, 17.9, 18.8, 20.8, and 24.5degrees 2θ, in combination with at least one peak selected from 5.4,9.4, 11.0, 12.3, 12.7, 14.2, 16.4, and 22.0 degrees 2θ, plus or minus0.2.

In one embodiment, Form 8 is characterized by XRPD peaks located at 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 5.4, 9.4, 11.0, 12.3, 12.7, 14.2, 16.0, 16.4, 17.9, 18.8,20.8, 22.0, and 24.5 degrees 2θ, plus or minus 0.2. In one embodiment,the solid form is characterized by 3 of the peaks. In one embodiment,the solid form is characterized by 5 of the peaks. In one embodiment,the solid form is characterized by 7 of the peaks. In one embodiment,the solid form is characterized by 9 of the peaks. In one embodiment,the solid form is characterized by 11 of the peaks. In one embodiment,the solid form is characterized by 13 of the peaks. In one embodiment,the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form 8 has an XRPD pattern substantially as shown inFIG. 20.

In one embodiment, Form 8 is an unsolvated solid form of Compound 1.

In one embodiment, Form 8 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at about 156° C.

All of the combinations of the above embodiments are encompassed by thisapplication.

In one embodiment, the diameter of the particle of the solid formsprovided herein (e.g., Form 1, Form 2, Form 3, Form 4, Form 5, Form 6,Form 7, or Form 8) is from about 0.1 μm to about 150 μm, from about 0.1μm to about 125 μm, from about 0.1 μm to about 100 μm, from about 0.1 μmto about 75 μm, from about 0.1 μm to about 50 μm, from about 1 μm toabout 50 μm, from about 0.1 μm to about 10 μm, from about 0.1 μm toabout 7 μm, or from about 0.5 μm to about 5 μm. In one embodiment, thediameter is from about 0.5 μm to about 5 μm. In another embodiment, thediameter is from about 0.6 μm to about 4.8 μm.

In one embodiment, provided herein is a composition comprising a solidform of a compound of formula (I), or a salt, or solvate, or solvate ofa salt thereof, or a mixture thereof, wherein the compound has a puritygreater than about 98.0% as determined by HPLC. In one embodiment, thecompound of formula (I) has a purity of about 98.5%, about 99.0%, about99.5%, about 99.6%, about 99.9%, or about 99.91%.

5.2.2. Process of Preparing Solid Forms of Compound 1

Provided herein is a process of preparing a compound of Formula (I),wherein the compound is polymorph Form 1 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 1 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 1 polymorph(s) into Form 1 of a compound of Formula (I); and

(ii) recovering said polymorph Form 1.

In one embodiment, the non-Form 1 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 1 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 1 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is an alcohol. In oneembodiment, the solvent is ethanol, 2-methoxyethanol, methanol,ethyleneglycol, or isopropyl alcohol. In one embodiment, the solvent isethyl acetate, methyl isobutyl ketone, toluene, 1,2-dimethoxyethane,N,N-dimethylformamide, acetonitrile, ethyleneglycol, anisole, or water.In one embodiment, the solvent is ethanol In one embodiment, the solventsystem comprises a mixture of two solvents. In one embodiment, thesolvent system is a mixture of two solvents. In one embodiment, themixture of two solvents is a mixture of anisole and isopropyl alcohol, amixture of anisole and ethanol, a mixture of anisole and toluene, amixture of acetonitrile and water, a mixture of toluene and ethanol, amixture of acetone and water, a mixture of isopropyl alcohol and water,a mixture of ethanol and water, a mixture of N,N-dimethylformamide andwater, a mixture of N,N-acetamide and water, a mixture ofdimethylsulfoxide and water, or a mixture of anisole and methanol In oneembodiment, the mixture of two solvents is a mixture of isopropylalcohol and water. In one embodiment, the volume ratio of isopropylalcohol to water is from about 1:4 to about 4:1. In one embodiment, thevolume ratio of isopropyl alcohol to water is about 1:1 or about 3:2. Inone embodiment, the volume ratio of isopropyl alcohol to water is about1:2. In one embodiment, the mixture of two solvents is a mixture ofacetone and water. In one embodiment, the mixture of two solvents is amixture of ethanol and water. In one embodiment, the mixture of twosolvents is a mixture of acetonitrile and water. In one embodiment, thevolume ratio of acetonitrile to water is from about 1:4 to about 8:1. Inone embodiment, the volume ratio of acetonitrile to water is about 4:1.In one embodiment, the volume ratio of acetonitrile to water is about2:3. In one embodiment, the solvent system comprises a mixture of threesolvents. In one embodiment, the mixture of three solvents is a mixtureof ethanol, water, and DCM.

In one embodiment, the non-Form 1 polymorph is amorphous compound ofFormula (I). In one embodiment, the non-Form 1 polymorph is Form 2 of acompound of Formula (I). In one embodiment, the period of timesufficient to convert at least about 50% of the total amount of non-Form1 polymorph(s) into Form 1 of a compound of Formula (I) is about 1 hr,about 2 hr, about 5 hr, about 10 hr, about 12 hr, about 20 hr, about 24hr, about 30 hr, about 40 hr, about 48 hr, or about 72 hr.

In one embodiment, the non-form 1 polymorph of a compound of Formula (I)is exposed to isopropyl alcohol and water, e.g., at a 1:1 volume ratio.Another volume of water is added at about 60° C., such that the finalvolume ratio of isopropyl alcohol to water is 1:2. The mixture is agedat about 60° C. for about 30 mins, 1 hr, about 2 hr, about 5 hr, about10 hr, about 12 hr, about 20 hr, about 24 hr, about 30 hr, about 40 hr,about 48 hr, or about 72 hr.

In one embodiment, the non-form 1 polymorph of a compound of Formula (I)is exposed to acetone and water, e.g., at a 4:1 volume ratio, at about50° C. to about 60° C. The solvent is exchanged from acetone/water toisopropyl alcohol to a final volume of about 30 volumes. The mixture isaged at about 60° C. for about 30 mins, 1 hr, about 2 hr, about 5 hr,about 10 hr, about 12 hr, about 14 hours, about 20 hr, about 24 hr,about 30 hr, about 40 hr, about 48 hr, or about 72 hr.

In one embodiment, Form 1 is prepared by crystallization orrecrystallization of a compound of Formula (I) from one or moresolvents. In one embodiment, the solvent is ethanol, 2-methoxyethanol,methanol, ethyleneglycol, or isopropyl alcohol. In one embodiment, thesolvent is ethyl acetate, methyl isobutyl ketone, toluene,1,2-dimethoxyethane, N,N-dimethylformamide, acetonitrile,ethyleneglycol, anisole, or water. In one embodiment, the solvent isethanol.

In one embodiment, Form 1 is prepared by crystallization orrecrystallization of a compound of Formula (I) from a solvent comprisingan alcohol. In one embodiment, the solvent is isopropyl alcohol. In oneembodiment, the solvent is ethanol.

In one embodiment, Form 1 is prepared by crystallization orrecrystallization of a compound of Formula (I) from a solvent comprisinga mixture of two solvents. In one embodiment, the mixture of twosolvents is a mixture of anisole and isopropyl alcohol, a mixture ofanisole and ethanol, a mixture of anisole and toluene, a mixture ofacetonitrile and water, a mixture of toluene and ethanol, a mixture ofacetone and water, a mixture of isopropyl alcohol and water, a mixtureof ethanol and water, a mixture of N,N-dimethylformamide and water, amixture of N,N-acetamide and water, a mixture of dimethylsulfoxide andwater, or a mixture of anisole and methanol In one embodiment, themixture of two solvents is a mixture of isopropyl alcohol and water. Inone embodiment, the volume ratio of isopropyl alcohol to water is fromabout 1:4 to about 4:1. In one embodiment, the volume ratio of isopropylalcohol to water is about 1:1 or about 3:2. In one embodiment, thevolume ratio of isopropyl alcohol to water is about 1:2. In oneembodiment, the mixture of two solvents is a mixture of acetone andwater. In one embodiment, the mixture of two solvents is a mixture ofethanol and water. In one embodiment, the mixture of two solvents is amixture of acetonitrile and water.

In one embodiment, Form 1 is prepared by crystallization orrecrystallization of a compound of Formula (I) from a solvent comprisinga mixture of an alcohol and water. In one embodiment, the solvent is amixture of from about 30% to about 90% alcohol in water. In oneembodiment, the solvent is a mixture of from about 40% to about 80%alcohol in water. In one embodiment, the solvent is a mixture ofisopropyl alcohol and water. In one embodiment, the solvent is a mixtureof about 3:2 isopropyl alcohol and water. In one embodiment, the solventis a mixture of ethanol and water. In one embodiment, the solvent is amixture of from about 20% to about 90% ethanol in water. In oneembodiment, the solvent is a mixture of from about 40% to about 80%ethanol in water. In one embodiment, the solvent is a mixture of about40% ethanol in water. In one embodiment, the solvent is a mixture ofabout 60% ethanol in water. In one embodiment, the solvent is a mixtureof about 80% ethanol in water. In one embodiment, the solvent furthercomprises DCM. In one embodiment, the crystallization orrecrystallization comprises one or more (e.g., 1, 2, 3, 4, 5 or 6)heating and cooling cycles. In one embodiment, the crystallization orrecrystallization comprises 3 heating and cooling cycles. In oneembodiment, the crystallization or recrystallization comprises 4 heatingand cooling cycles. In one embodiment, the heating phase comprisesheating at from about 50° C. to about 60° C. for a period of time (e.g.,from about 1 hour to about 6 hours, e.g., about 3 hours). In oneembodiment, the cooling phase comprising holding at room temperature fora period of time (e.g., from about 1 hour to about 6 hours, e.g., about2 hours).

In one embodiment, Form 1 is prepared by crystallization orrecrystallization of a compound of Formula (I) from a solvent comprisinga mixture of acetonitrile and water. In one embodiment, the volume ratioof acetonitrile to water is from about 1:4 to about 8:1. In oneembodiment, the volume ratio of acetonitrile to water is about 4:1. Inone embodiment, the volume ratio of acetonitrile to water is about 2:3.

Provided herein is a process of preparing a compound of Formula (I),wherein the compound is polymorph Form 2 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 2 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 2 polymorph(s) into Form 2 of a compound of Formula (I); and

(ii) recovering said polymorph Form 2.

In one embodiment, the non-Form 2 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 2 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 2 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is dichloromethane,acetone, tetrahydrofuran, water, 1-propanol, or chloroform. In oneembodiment, the mixture of two solvents is a mixture of dichloromethaneand acetone, a mixture of tetrahydrofuran and water, a mixture ofdichloromethane and ethanol, or a mixture of dichloromethane andmethanol In one embodiment, the mixture of two solvents is a mixture ofdichloromethane and acetone. In one embodiment, the non-Form 2 polymorphof a compound of Formula (I) is exposed to a mixture of three solvents.In one embodiment, the mixture of three solvents is a mixture ofdichloromethane, ethanol, and water. In one embodiment, the non-Form 2polymorph is amorphous compound of Formula (I). In one embodiment, theperiod of time sufficient to convert at least about 50% of the totalamount of non-Form 2 polymorph(s) into Form 2 of a compound of Formula(I) is about 1 hr, about 2 hr, about 5 hr, about 10 hr, about 12 hr,about 20 hr, about 24 hr, about 30 hr, about 40 hr, about 48 hr, orabout 72 hr.

In one embodiment, Form 2 is obtained from maturation e.g., in1-propanol, acetone or dichloromethane, or a mixture of dichloromethaneand acetone or a mixture of tetrahydrofuran and water. In oneembodiment, Form 2 is obtained from dichloromethane at about 5° C.

Provided herein is a process of preparing a compound of Formula (I),wherein the compound is polymorph Form 3 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 3 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 3 polymorph(s) into Form 3 of a compound of Formula (I); and

(ii) recovering said polymorph Form 3.

In one embodiment, the non-Form 3 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 3 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 3 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is methyl ethyl ketone,tert-butylmethyl ether, 2-methyl-1-propanol, 2-methyltetrahydrofuran,isopropyl alcohol, ethanol, toluene, 1-propanol, acetone, oracetonitrile. In one embodiment, the mixture of two solvents is amixture of 2-methyltetrahydrofuran and isopropyl alcohol, a mixture of2-methyltetrahydrofuran and ethanol, a mixture of2-methyltetrahydrofuran and toluene, or a mixture of acetonitrile andwater. In one embodiment, the non-Form 3 polymorph is amorphous compoundof Formula (I). In one embodiment, the period of time sufficient toconvert at least about 50% of the total amount of non-Form 3polymorph(s) into Form 3 of a compound of Formula (I) is about 1 hr,about 2 hr, about 5 hr, about 10 hr, about 12 hr, about 20 hr, about 24hr, about 30 hr, about 40 hr, about 48 hr, or about 72 hr.

In one embodiment, Form 3 is obtained from maturation in one solvent ora mixture of one or more solvents. In one embodiment, Form 3 is obtainedat about 5° C.

Provided herein is a process of preparing a compound of Formula (I),wherein the compound is polymorph Form 4 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 4 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 4 polymorph(s) into Form 4 of a compound of Formula (I); and

(ii) recovering said polymorph Form 4.

In one embodiment, the non-Form 4 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 4 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 4 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is 1-propanol, acetone,2-methyl-1-propanol, 1,4-dioxane, chloroform, tetrahydrofuran,2-methoxyethanol, isopropyl alcohol, water, anisole, toluene, ordimethylsulfoxide. In one embodiment, the mixture of two solvents is amixture of anisole and tetrahydrofuran, a mixture of toluene andtetrahydrofuran, a mixture of toluene and isopropyl alcohol, or amixture of isopropyl alcohol and water. In one embodiment, the non-Form4 polymorph is amorphous compound of Formula (I). In one embodiment, theperiod of time sufficient to convert at least about 50% of the totalamount of non-Form 4 polymorph(s) into Form 4 of a compound of Formula(I) is about 1 hr, about 2 hr, about 5 hr, about 10 hr, about 12 hr,about 20 hr, about 24 hr, about 30 hr, about 40 hr, about 48 hr, orabout 72 hr.

In one embodiment, Form 4 is obtained from maturation in one solvent ora mixture of one or more solvents. In one embodiment, Form 4 is obtainedat about 5° C.

Provided herein is a process of preparing a compound of Formula (I),wherein the compound is polymorph Form 5 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 5 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 5 polymorph(s) into Form 5 of a compound of Formula (I); and

(ii) recovering said polymorph Form 5.

In one embodiment, the non-Form 5 polymorph of a compound of Formula (I)is exposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is anisole. In oneembodiment, the non-Form 5 polymorph is amorphous compound of Formula(I). In one embodiment, the period of time sufficient to convert atleast about 50% of the total amount of non-Form 5 polymorph(s) into Form5 of a compound of Formula (I) is about 1 hr, about 2 hr, about 5 hr,about 10 hr, about 12 hr, about 20 hr, about 24 hr, about 30 hr, about40 hr, about 48 hr, or about 72 hr.

In one embodiment, Form 5 is obtained from maturation in one solvent ora mixture of one or more solvents. In one embodiment, Form 5 is obtainedat about 5° C.

Provided herein is a process of preparing compound of Formula (I),wherein the compound is polymorph Form 6 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 6 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 6 polymorph(s) into Form 6 of a compound of Formula (I); and

(ii) recovering said polymorph Form 6.

In one embodiment, the non-Form 6 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 6 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 6 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is nitromethane,acetonitrile, or water. In one embodiment, the solvent is nitromethaneIn one embodiment, the mixture of two solvents is a mixture ofnitromethane and water or a mixture of acetonitrile and water. In oneembodiment, the mixture of two solvents is a mixture of acetonitrile andwater. In one embodiment, the volume ratio of acetonitrile to water is1:1. In one embodiment, the non-Form 6 polymorph is amorphous compoundof Formula (I). In one embodiment, the period of time sufficient toconvert at least about 50% of the total amount of non-Form 6polymorph(s) into Form 6 of a compound of Formula (I) is about 1 hr,about 2 hr, about 5 hr, about 10 hr, about 12 hr, about 20 hr, about 24hr, about 30 hr, about 40 hr, about 48 hr, or about 72 hr.

In one embodiment, Form 6 is obtained from maturation in one solvent ora mixture of one or more solvents. In one embodiment, Form 6 is obtainedat about 5° C.

Provided herein is a process of preparing compound of Formula (I),wherein the compound is polymorph Form 7 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 7 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to one or more solvent for a periodof time sufficient to convert at least about 50% of the total amount ofnon-Form 7 polymorph(s) into Form 7 of a compound of Formula (I); and

(ii) recovering said polymorph Form 7.

In one embodiment, the non-Form 7 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 7 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 7 polymorph of a compound of Formula (I) isexposed to one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is methyl ethyl ketone,1-propanol, acetone, or tert-butyl methyl ether. In one embodiment, thenon-Form 7 polymorph is amorphous compound of Formula (I). In oneembodiment, the period of time sufficient to convert at least about 50%of the total amount of non-Form 7 polymorph(s) into Form 7 of a compoundof Formula (I) is about 1 hr, about 2 hr, about 5 hr, about 10 hr, about12 hr, about 20 hr, about 24 hr, about 30 hr, about 40 hr, about 48 hr,or about 72 hr.

Provided herein is a process of preparing compound of Formula (I),wherein the compound is polymorph Form 8 of a compound of Formula (I),or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof; and the process comprises:

(i) exposing a composition comprising at least one non-Form 8 polymorphof a compound of Formula (I), or a salt, or solvate, or solvate of asalt thereof, or a mixture thereof, to heat for a period of timesufficient to convert at least about 50% of the total amount of non-Form8 polymorph(s) into Form 7 of a compound of Formula (I); and

(ii) recovering said polymorph Form 8.

In one embodiment, the non-Form 8 polymorph is Form 6 compound ofFormula (I). In one embodiment, the period of time sufficient to convertat least about 50% of the total amount of non-Form 8 polymorph(s) intoForm 8 of a compound of Formula (I) is about 1 hr, about 2 hr, about 5hr, about 10 hr, about 12 hr, about 20 hr, about 24 hr, about 30 hr,about 40 hr, about 48 hr, or about 72 hr.

5.2.3. Solid Forms Comprising Compound 1 and a Coformer

In certain embodiments, the solid forms provided herein further comprisea coformer. In certain embodiments, provided herein is a solid formcomprising a compound of Formula (I):

or a salt, or solvate (e.g., hydrate), or solvate of a salt thereof, ora mixture thereof, and a coformer.

In one embodiment, provided herein a solid form comprising a free baseof Compound 1, or a solvate (e.g., hydrate) thereof, and a coformer. Inone embodiment, provided herein is an unsolvated solid form comprisingCompound 1 and a coformer. In one embodiment, provided herein is ananhydrous solid form comprising Compound 1 and a coformer. In oneembodiment, provided herein is a solvated solid form comprising Compound1 and a coformer. In one embodiment, provided herein is a hydrate solidform comprising Compound 1 and a coformer.

It is contemplated that Compound 1, or a salt, or solvate (e.g.,hydrate), or solvate of a salt thereof, or a mixture thereof, and acoformer can exist in a variety of solid forms. Such solid forms includecrystalline solids or mixtures of crystalline and amorphous solids. Inone embodiment, the solid form is substantially crystalline. In oneembodiment, the solid form is crystalline. In one embodiment, the solidform is a cocrystal.

In some embodiments, the molar ratio of Compound 1 to the solvent/waterin the solid form ranges from about 10:1 to about 1:10. In someembodiments, the molar ratio of Compound 1 to the solvent/water in thesolid form ranges from about 5:1 to about 1:5. In some embodiments, themolar ratio of Compound 1 to the solvent/water in the solid form rangesfrom about 3:1 to about 1:3. In some embodiments, the molar ratio ofCompound 1 to the solvent/water in the solid form ranges from about 2:1to about 1:2. In one embodiment, the molar ratio is about 1:2 (i.e.,bis-solvate/hydrate). In another embodiment, the molar ratio is about1:1 (i.e., mono-solvate/hydrate). In yet another embodiment, the molarratio is about 2:1 (i.e., hemi-solvate/hydrate).

The ratio of Compound 1 to coformer may be stoichiometric ornon-stoichiometric. In one embodiment, the ratio of Compound 1 tocoformer ranges from about 5:1 to about 1:5. In one embodiment, theratio of Compound 1 to coformer is about 5:1, 4:1, 3:1, 2.5:1, 2:1,1.5:1, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, or 1:5. In one embodiment, theratio of Compound 1 to coformer is about 1:1. In one embodiment, theco-crystal comprises more than one coformers. In one embodiment, theco-crystal comprises two coformers.

In one embodiment, the coformer is one or more of citric acid, L-malicacid, L-tartaric acid, fumaric acid, succinic acid, maleic acid, sorbicacid, ketoglutaric acid, salicylic acid, benzoic acid, 3-hydroxybenzoicacid, 2,4-dihydroxybenzoic acid, 4-aminobenzoic acid, orotic acid, urea,nicotinic acid, isonicotinic acid, nicotinamide, isonicotinamide,saccharin, L-lactic acid, L-serine, L-proline, glycine, maltol,succinimide, sulfacetamide, and p-toluenesulfonic acid monohydrate.

In one embodiment, the coformer is L-tartaric acid. In anotherembodiment, the coformer is salicylic acid.

5.2.3.1 Form P1C3 of a Solid Form Comprising Compound 1 and L-tartaricAcid

In some embodiments, provided herein is Form P1C3 of a solid formcomprising Compound 1 and L-tartaric acid. In one embodiment, Form P1C3is a crystalline hydrate solid form comprising Compound 1 and L-tartaricacid. In some embodiments, Form P1C3 is substantially free of amorphousCompound 1. In some embodiments, Form P1C3 of Compound 1 issubstantially free of other crystalline forms (i.e., polymorphs) ofCompound 1. In some embodiments, Form P1C3 is provided as substantiallypure Form P1C3.

In one embodiment, the molar ratio of Compound 1 to L-tartaric acid inForm P1C3 ranges from about 1:2 to 2:1. In one embodiment, the molarratio of Compound 1 to L-tartaric acid in Form P1C3 is about 1:1. In oneembodiment, Form P1C3 further comprises water. In one embodiment, themolar ratio of Compound 1 to water in Form P1C3 ranges from about 1:2 to2:1. In one embodiment, the molar ratio of Compound 1 to water in FormP1C3 is about 1:1. In one embodiment, the molar ratio of Compound1:L-tartaric acid:water in Form P1C3 is about 1:1:1.

A representative XRPD pattern of Form P1C3 is provided in FIG. 22.

In one embodiment, Form P1C3 has an XRPD pattern comprising peaks at11.2, 17.4, and 17.7 degrees 2θ, plus or minus 0.2. In one embodiment,Form P1C3 has an XRPD pattern further comprising at least one peakselected from 21.2 and 22.5 degrees 2θ, plus or minus 0.2. In oneembodiment, Form P1C3 has an XRPD pattern comprising peaks at 11.2,17.4, 17.7, 21.2, and 22.5 degrees 2θ, in combination with at least onepeak selected from 10.7, 11.6, 17.0, 20.6, 20.8, 21.4, 22.2, 23.2, 23.6,and 24.2 degrees 2θ, plus or minus 0.2.

In one embodiment, Form P1C3 is characterized by XRPD peaks located at1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 10.7, 11.2, 11.6, 17.0, 17.4, 17.7, 20.6, 20.8, 21.2, 21.4,22.2, 22.5, 23.2, 23.6, and 24.2 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form P1C3 has an XRPD pattern substantially as shownin FIG. 22.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form P1C3 ofCompound 1 is provided in FIG. 23.

In one embodiment, Form P1C3 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at about 129° C. and/ora peak temperature at about 149° C. In one embodiment, Form P1C3 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 23.

In one embodiment, Form P1C3 exhibits a weight loss of about 0.5% uponheating from about 30° C. to about 100° C., a weight loss of about 1.9%upon heating from about 100° C. to about 160° C., and a weight loss ofabout 15.9% upon heating from about 170° C. to about 260° C. In oneembodiment, Form P1C3 is characterized by a TGA thermogram substantiallyas shown in the TGA thermogram presented in FIG. 23.

A representative gravimetric vapor sorption (GVS) isotherm of Form P1C3is presented in FIG. 28. In one embodiment, Form P1C3 exhibits a weightincrease of about 3.7% when subjected to an increase in relativehumidity from about 0 to about 90% relative humidity. In one embodiment,Form P1C3 is characterized by a GVS thermogram substantially as shown inthe GVS thermogram presented in FIG. 28.

In one embodiment, preparation of Form P1C3 comprises grinding a mixtureof Compound 1 and L-tartaric acid in the presence of a solvent. In oneembodiment, the solvent is nitromethane In one embodiment, thenitromethane is not anhydrous, i.e., contains certain amount (e.g.,about 5%) of water. In one embodiment, preparation of Form P1C3comprises grinding a 1:1 mixture of Compound 1 and L-tartaric acid inthe presence of nitromethane.

In one embodiment, preparation of Form P1C3 comprises slow cooling asolution of Compound 1 and L-tartaric acid in a solvent. In oneembodiment, the solvent is nitromethane In one embodiment, thenitromethane is not anhydrous, i.e., contains certain amount (e.g.,about 5%) of water. In one embodiment, preparation of Form P1C3comprises slow cooling a solution of 1:1 Compound 1 and L-tartaric acidin nitromethane with about 5% water. In one embodiment, the solution iscooled from about 50° C. to about 5° C. at a rate of from about 0.1 toabout 0.25° C./min.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.3.2 Form P1C9 of a Solid Form Comprising Compound 1 and SalicylicAcid

In some embodiments, provided herein is Form P1C9 of a solid formcomprising Compound 1 and salicylic acid. In one embodiment, Form P1C9is a crystalline hydrate solid form comprising Compound 1 and salicylicacid. In some embodiments, Form P1C9 is substantially free of amorphousCompound 1. In some embodiments, Form P1C9 of Compound 1 issubstantially free of other crystalline forms (i.e., polymorphs) ofCompound 1. In some embodiments, Form P1C9 is provided as substantiallypure Form P1C9.

In one embodiment, the molar ratio of Compound 1 to salicylic acid inForm P1C9 ranges from about 1:1 to 3:1. In one embodiment, the molarratio of Compound 1 to salicylic acid in Form P1C9 is about 2:1. In oneembodiment, Form P1C9 further comprises water. In one embodiment, themolar ratio of Compound 1 to water in Form P1C9 ranges from about 1:3 to1:5. In one embodiment, the molar ratio of Compound 1 to water in FormP1C9 is about 1:4. In one embodiment, the molar ratio of Compound1:salicylic acid:water in Form P1C9 is about 1:0.5:4.

A representative XRPD pattern of Form P1C9 is provided in FIG. 24.

In one embodiment, Form P1C9 has an XRPD pattern comprising peaks at6.9, 10.1, and 12.0 degrees 2θ, plus or minus 0.2. In one embodiment,Form P1C9 has an XRPD pattern further comprising at least one peakselected from 17.8 and 20.0 degrees 2θ, plus or minus 0.2. In oneembodiment, Form P1C9 has an XRPD pattern comprising peaks at 6.9, 10.1,12.0, 17.8, and 20.0 degrees 2θ, in combination with at least one peakselected from 4.7, 6.0, 12.7, 13.7, 15.0, 16.2, 24.2, 24.6, 26.1, and28.3 degrees 2θ, plus or minus 0.2.

In one embodiment, Form P1C9 is characterized by XRPD peaks located at1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 4.7, 6.0, 6.9, 10.1, 12.0, 12.7, 13.7, 15.0, 16.2, 17.8,20.0, 24.2, 24.6, 26.1, and 28.3 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form P1C9 has an XRPD pattern substantially as shownin FIG. 24.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form P1C9 ofCompound 1 is provided in FIG. 25.

In one embodiment, Form P1C9 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 43° C. and/or apeak temperature at about 75° C., or an endothermic event with an onsettemperature at about 120° C. and/or a peak temperature at about 127° C.In one embodiment, Form P1C9 exhibits, as characterized by DSC, anendothermic event with an onset temperature at about 43° C. and/or apeak temperature at about 75° C., and an endothermic event with an onsettemperature at about 120° C. and/or a peak temperature at about 127° C.In one embodiment, Form P1C9 is characterized by a DSC thermogramsubstantially as shown in the DSC thermogram presented in FIG. 25.

In one embodiment, Form P1C9 exhibits a weight loss of about 10.15% uponheating from about 30° C. to about 100° C. In one embodiment, Form P1C9is characterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 25.

In one embodiment, preparation of Form P1C9 comprises sonicating amixture of Compound 1 and salicylic acid in the presence of a solvent.In one embodiment, the solvent is a mixture of acetonitrile and water.In one embodiment, the solvent is a 1:1 mixture of acetonitrile andwater. In one embodiment, preparation of Form P1C9 comprises sonicatinga 1:1 mixture of Compound 1 and salicylic acid in the presence of a 1:1mixture of acetonitrile and water. In one embodiment, the preparationfurther comprises settling the material from sonicating step for aperiod of time. In one embodiment, the settling period is less thanabout 2 hours. In one embodiment, the settling period is about 30minutes.

All of the combinations of the above embodiments are encompassed by thisapplication.

5.2.3.3 Form P2C9 of a Solid Form Comprising Compound 1 and SalicylicAcid

In some embodiments, provided herein is Form P2C9 of a solid formcomprising Compound 1 and salicylic acid. In one embodiment, Form P2C9is a crystalline solvate solid form comprising Compound 1 and salicylicacid. In one embodiment, Form P2C9 is a crystalline acetonitrile solvatesolid form comprising Compound 1 and salicylic acid. In someembodiments, Form P2C9 is substantially free of amorphous Compound 1. Insome embodiments, Form P2C9 of Compound 1 is substantially free of othercrystalline forms (i.e., polymorphs) of Compound 1. In some embodiments,Form P2C9 is provided as substantially pure Form P2C9.

In one embodiment, the molar ratio of Compound 1 to salicylic acid inForm P2C9 ranges from about 1:2 to 2:1. In one embodiment, the molarratio of Compound 1 to salicylic acid in Form P2C9 is about 1:1. In oneembodiment, Form P2C9 further comprises acetonitrile. In one embodiment,the molar ratio of Compound 1 to acetonitrile in Form P2C9 ranges fromabout 1:1 to 3:1. In one embodiment, the molar ratio of Compound 1 toacetonitrle in Form P2C9 is about 1:0.5. In one embodiment, the molarratio of Compound 1:salicylic acid:acetonitrile in Form P2C9 is about1:1:0.5.

A representative XRPD pattern of Form P2C9 is provided in FIG. 26.

In one embodiment, Form P2C9 has an XRPD pattern comprising peaks at11.4, 13.4, and 24.0 degrees 2θ, plus or minus 0.2. In one embodiment,Form P2C9 has an XRPD pattern further comprising at least one peakselected from 25.1 and 26.9 degrees 2θ, plus or minus 0.2. In oneembodiment, Form P2C9 has an XRPD pattern comprising peaks at 11.4,13.4, 24.0, 25.1, and 26.9 degrees 2θ, in combination with at least onepeak selected from 8.5, 12.7, 16.0, 16.8, 18.7, 19.9, 21.7, 23.6, 28.3,and 28.7 degrees 2θ, plus or minus 0.2.

In one embodiment, Form P2C9 is characterized by XRPD peaks located at1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of the followingpositions: 8.5, 11.4, 12.7, 13.4, 16.0, 16.8, 18.7, 19.9, 21.7, 23.6,24.0, 25.1, 26.9, 28.3, and 28.7 degrees 2θ, plus or minus 0.2. In oneembodiment, the solid form is characterized by 3 of the peaks. In oneembodiment, the solid form is characterized by 5 of the peaks. In oneembodiment, the solid form is characterized by 7 of the peaks. In oneembodiment, the solid form is characterized by 9 of the peaks. In oneembodiment, the solid form is characterized by 11 of the peaks. In oneembodiment, the solid form is characterized by 13 of the peaks. In oneembodiment, the solid form is characterized by all of the peaks.

In some embodiments, the XRPD peaks above (degrees 2θ peaks) areobserved when analyzed using copper Kα radiation.

In one embodiment, Form P2C9 has an XRPD pattern substantially as shownin FIG. 26.

A representative overlay of thermal gravimetric analysis (TGA) anddifferential scanning calorimetry (DSC) thermograms for Form P2C9 ofCompound 1 is provided in FIG. 27.

In one embodiment, Form P2C9 exhibits an endothermic event, ascharacterized by DSC, with an onset temperature at about 78° C. and/or apeak temperature at about 96° C. In one embodiment, Form P2C9 ischaracterized by a DSC thermogram substantially as shown in the DSCthermogram presented in FIG. 27.

In one embodiment, Form P2C9 exhibits a weight loss of about 5.6% uponheating from about 30° C. to about 130° C. In one embodiment, Form P2C9is characterized by a TGA thermogram substantially as shown in the TGAthermogram presented in FIG. 27.

In one embodiment, preparation of Form P2C9 comprises sonicating amixture of Compound 1 and salicylic acid in the presence of a solvent.In one embodiment, the solvent is a mixture of acetonitrile and water.In one embodiment, the solvent is a 1:1 mixture of acetonitrile andwater. In one embodiment, preparation of Form P2C9 comprises sonicatinga 1:1 mixture of Compound 1 and salicylic acid in the presence of a 1:1mixture of acetonitrile and water. In one embodiment, the preparationfurther comprises settling the material from sonicating step for aperiod of time. In one embodiment, the settling period is at least about2 hours.

All of the combinations of the above embodiments are encompassed by thisapplication.

In one embodiment, the diameter of the particle of the solid formsprovided herein (e.g., Form P1C9, Form P1C9, or Form P2C9), wherein thediameter of the particle of the compound is from about 0.1 μm to about150 μm, from about 0.1 μm to about 125 μm, from about 0.1 μm to about100 μm, from about 0.1 μm to about 75 μm, from about 0.1 μm to about 50μm, from about 1 μm to about 50 μm, from about 1 μm to about 50 μm, fromabout 0.1 μm to about 10 μm, from about 0.1 μm to about 7 μm, or fromabout 0.5 μm to about 5 μm. In one embodiment, the diameter is fromabout 0.5 μm to about 5 μm. In one embodiment, the diameter is fromabout 0.6 μm to about 4.8 μm.

In one embodiment, provided herein is a composition comprising a solidform of a compound of formula (I), or a salt, or solvate, or solvate ofa salt thereof, or a mixture thereof, wherein the compound has a puritygreater than about 98.0% as determined by HPLC. In one embodiment, thecompound of formula (I) has a purity of about 98.5%, about 99.0%, about99.5%, about 99.6%, about 99.9%, or about 99.91%.

5.2.4 Methods for Analyzing Material

In some embodiments, provided herein are also methods for analyzing amaterial for the presence or amount of a solid form provided herein,comprising providing a material comprising a compound of formula (I), ora salt, solvate (e.g., hydrate), or solvate of a salt thereof, or amixture thereof; and using a characterization method to determinewhether a signatory characteristic associated with the solid form ispresent in the material by comparing the characteristic obtained fromthe material with a reference signatory characteristic; wherein theexistence of a characteristic substantially identical to the referencesignatory characteristic indicates the presence of the solid form in thematerial.

In one embodiment, the method further comprises selecting a batch as aresult of the determination based upon comparison to the referencestandard. In one embodiment, the method further comprises making adetermination regarding the quality of the material. In one embodiment,the method further comprises making a determination whether to use thematerial in the manufacturing of a pharmaceutical composition. In oneembodiment, the method further comprises making a determination whetherto use the material for treating a PI3K mediated disorder.

In one embodiment, the characterization method is one or more of XRPD,TGA, DSC, GVS, FT-IR, or NMR.

5.2.5. Process for Preparation of Amorphous Form of Compound 1

Provided herein is a process of preparing amorphous form of a compoundof Formula (I), wherein the amorphous form is made via a crystallineform. In one embodiment, provided herein is a process of preparingamorphous form of compound of Formula (I), wherein the amorphous form ismade via polymorph Form 1 of a compound of Formula (I), or a salt, orsolvate, or solvate of a salt thereof, or a mixture thereof.

In one embodiment, the process comprises:

(i) dissolving a solid form comprising a polymorphic form of thecompound of formula (I), or a salt, or solvate, or solvate of a saltthereof, or a mixture thereof, in one or more solvents to form asolution; and

(ii) removing the solvent of the solution to provide the amorphous formof the compound of formula (I).

In one embodiment, the polymorphic form is polymorph Form 1.

In one embodiment, the solid form further comprising amorphous form ofthe compound of formula (I).

In one embodiment, the process comprises:

(i) dissolving a solid form comprising polymorph Form 1 of the compoundof formula (I), or a salt, or solvate, or solvate of a salt thereof, ora mixture thereof, in one or more solvents to form a solution; and

(ii) removing the solvent of the solution to provide the amorphous formof the compound of formula (I).

In one embodiment, the process comprises:

(i) exposing a composition comprising at least one non-Form 1 polymorphor amorphous form of a compound of Formula (I), or a salt, or solvate,or solvate of a salt thereof, or a mixture thereof, to one or moresolvent for a period of time sufficient to convert at least about 50% ofthe total amount of non-Form 1 polymorph(s) into Form 1 of a compound ofFormula (I);

(ii) recovering said polymorph Form 1;

(iii) dissolving said polymorph Form 1 in one or more solvents to form asolution; and

(iv) removing the solvent of the solution to provide the amorphous formof the compound of formula (I).

In one embodiment, the solvent of the solution is removed bylyophilization. In another embodiment, the solvent of the solution isremoved by spray drying.

In one embodiment, the non-Form 1 polymorph of a compound of Formula (I)is exposed to one solvent. In one embodiment, the non-Form 1 polymorphof a compound of Formula (I) is exposed to a mixture of two solvents. Inone embodiment, the non-Form 1 polymorph of a compound of Formula (I) isexposed to a one or more solvents. In one embodiment, the solvent is anorganic solvent. In one embodiment, the solvent is an alcohol. In oneembodiment, the solvent is ethanol, 2-methoxyethanol, methanol,ethyleneglycol, or isopropyl alcohol. In one embodiment, the solvent isethyl acetate, methyl isobutyl ketone, toluene, 1,2-dimethoxyethane,N,N-dimethylformamide, acetonitrile, ethyleneglycol, anisole, or water.In one embodiment, the solvent is ethanol In one embodiment, the mixtureof two solvents is a mixture of anisole and isopropyl alcohol, a mixtureof anisole and ethanol, a mixture of anisole and toluene, a mixture ofacetonitrile and water, a mixture of toluene and ethanol, a mixture ofacetone and water, or a mixture of isopropyl alcohol and water. In oneembodiment, the mixture of two solvents is a mixture of isopropylalcohol and water. In one embodiment, the volume ratio of isopropylalcohol to water is 1:1 or 3:2. In one embodiment, the volume ratio ofisopropyl alcohol to water is 1:2. In one embodiment, the mixture of twosolvents is a mixture of acetone and water. In one embodiment, thenon-Form 1 polymorph is amorphous compound of Formula (I). In oneembodiment, the period of time sufficient to convert at least about 50%of the total amount of non-Form 1 polymorph(s) into Form 1 of a compoundof Formula (I) is about 1 hr, about 2 hr, about 5 hr, about 10 hr, about12 hr, about 20 hr, about 24 hr, about 30 hr, about 40 hr, about 48 hr,or about 72 hr.

In one embodiment, the non-form 1 polymorph of a compound of Formula (I)is exposed to isopropyl alcohol and water, e.g., at a 1:1 volume ratio.Another volume of water is added at about 60° C., such that the finalvolume ratio of isopropyl alcohol to water is 1:2. The mixture is agedat about 60° C. for about 30 mins, 1 hr, about 2 hr, about 5 hr, about10 hr, about 12 hr, about 20 hr, about 24 hr, about 30 hr, about 40 hr,about 48 hr, or about 72 hr.

In one embodiment, the non-form 1 polymorph of a compound of Formula (I)is exposed to acetone and water, e.g., at a 4:1 volume ratio, at about50° C. to about 60° C. The solvent is exchanged from acetone/water toisopropyl alcohol to a final volume of about 30 volumes. The mixture isaged at about 60° C. for about 30 mins, 1 hr, about 2 hr, about 5 hr,about 10 hr, about 12 hr, about 14 hours, about 20 hr, about 24 hr,about 30 hr, about 40 hr, about 48 hr, or about 72 hr.

In one embodiment, the solvent in which compound of Formula (I) isdissolved in is DCM, an alcohol, or a mixture thereof. In oneembodiment, the alcohol is MeOH. In one embodiment, the alcohol is2-propanol. In another embodiment, a polymer is added prior to spraydrying. In another embodiment, the polymer is PVP/VA 64. In anotherembodiment, the polymer is HPMC-AS.

5.3 Process for Preparation

In certain embodiments, provided herein is a process of preparing acompound of Formula (I):

or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof,comprising coupling compound C of formula:

with a carboxylic acid of Formula G:

to provide the compound of Formula (I).

In one embodiment, the coupling occurs in the presence of a couplingreagent. In one embodiment, the coupling reagent is a carbodiimide, atriazine, a phosphonium, an uronium, or a mixed anhydride, or a mixturethereof. In one embodiment, the coupling reagent isN,N′-dicyclohexylcarbodimide (DCC), N,N′-diisopropylcarbodiimide (DIC),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI),hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole (HOAt),2-propanephosphonic acid anhydride (T3P),1-[(dimethylamino)(morpholino)methylene]-1H-[1,2,3]triazolo[4,5-b]pyridine-1-ium3-oxide hexafluorophosphate (HDMA),N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uroniumhexafluorophosphate (HBTU),(1-cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate (COMU), benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP),(benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate(PyBOP),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU), diethyl phosphorocyanidate (DECP),diethyl phosphorochloridate (DEPC), diphenyl phosphorazidate (DPPA)phosphoric acid bis(2-oxazolidide) chloride (BOPCl),chlorodimethoxytriazine or its N-methylmopholinium adduct,3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT), bromotris(dimethylamino) phosphonium hexafluorophosphate) (BroP),(EtO)₂P(O)—Cl, (EtO)₂P(O)-Oxyma, pivaloyl chloride, iso-butylchloroformate, 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT), or4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride(DMTMM) or its BF₄ analog, or a mixture thereof. In one embodiment, thecoupling reagent is EDCI. In one embodiment, the coupling reagent isDMTMM.

In one embodiment, the coupling occurs in the presence of an activator.In one embodiment, the activator is HOBt, HBTriazinone, ethyl2-cyano-2-(hydroxyimino)acetate (Oxyma), NHS, or ethyl(hydroxyimino)cyanoacetate potassium salt (K-Oxyma). In one embodiment,the activator is HOBt.

In one embodiment, the coupling occurs in the presence of a base. In oneembodiment, the base is Et₃N, DIPEA, pyridine, NMM, DBU, NaOH, or DMAP.In one embodiment, the base is Et₃N. In one embodiment, the base isDIPEA.

In one embodiment, the coupling occurs in the presence of a solvent. Inone embodiment, the solvent is DMF, NMP, acetonitrile, EtOH, acetone,DCM, MeOH, or water, or a mixture thereof.

In one embodiment, the coupling occurs in the presence of a carbodiimidecoupling reagent. In one embodiment, the carbodiimide is1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. In one embodiment, thecoupling occurs in the presence of hydroxybenzotriazole (HOBt). In oneembodiment, the coupling occurs in the presence of a base. In oneembodiment, the base is DIPEA. In one embodiment, the DIPEA is in DMF.In one embodiment, the coupling occurs under an inert atmosphere.

In one embodiment, the coupling occurs in the presence of a triazinecoupling reagent. In one embodiment, the triazine is DMTMM. In oneembodiment, the coupling occurs in the presence of a base. In oneembodiment, the base is Et₃N, DIPEA, pyridine, NMM, DBU, NaOH, or DMAP.In one embodiment, the base is Et₃N. In one embodiment, the couplingoccurs in a solvent of acetonitrile, EtOH, acetone, DCM, MeOH, or water,or a mixture thereof. In one embodiment, the solvent is a mixture ofacetonitrile and water (e.g., 4:1 v/v), a mixture of EtOH and water(e.g., 3:1 v/v), a mixture of EtOH, water, and DCM (e.g., 14.4:4.8:1v/v/v), a mixture of acetone and water (e.g., 4:1 v/v), a mixture of DCMand MeOH (e.g., 4:1 v/v), a mixture of DCM and EtOH (e.g., 4:1 v/v), orDCM. In one embodiment, the solvent is a mixture of acetonitrile andwater. In one embodiment, the volume ratio of acetonitrile to water isabout 4:1.

In one embodiment, the coupling occurs in the presence of T3P and DIPEAin DMF.

In certain embodiments, provided herein is a process of preparing acompound of Formula (I):

or a salt, or solvate, or solvate of a salt thereof, or a mixturethereof,comprising coupling compound C of formula:

with an ester of Formula D:

to provide the compound of Formula (I).

In one embodiment, the coupling occurs in the presence of one or moresolvents. In another embodiment, the coupling occurs in the presence ofa base and one or more solvents. In one embodiment, the base is anamine. In one embodiment, the amine is N,N-diisopropylethylamine(DIPEA). In one embodiment, the solvent is an organic solvent. In oneembodiment, the organic solvent is selected from acetonitrile,dimethylformamide, tetrahydrofuran, 2-methyltetrahydrofuran, anddichloromethane, or a mixture thereof. In one embodiment, the organicsolvent is acetonitrile. In one embodiment, the organic solvent is amixture of DCM and ethanol In one embodiment, the volume ratio of DCM toethanol is from about 8:1 to about 2:1. In one embodiment, the volumeratio of DCM to ethanol is about 4:1. In one embodiment, the couplingoccurs in the presence of a mixture of two solvents. In anotherembodiment, the mixture of solvents is water and acetonitrile. In oneembodiment, the volume ratio of water to acetonitrile is about 1:4. Inone embodiment, the coupling occurs at a temperature from about 30° C.to about 80° C., from about 40° C. to about 70° C., or from about 55° C.to about 65° C. In one embodiment, the temperature is about 60° C.

In certain embodiments, provided herein is a process of preparingcompound C of formula:

comprising coupling compound A of formula:

with an alkyne of Formula E:

In one embodiment, the coupling occurs in the presence of a catalyst, aligand, or a catalyst/ligand complex; a base; and a solvent.

In one embodiment, the catalyst is a palladium (Pd) catalyst, a nickel(Ni) catalyst, a copper (Cu) catalyst, or a mixture thereof. In oneembodiment, the catalyst is a Pd catalyst. In one embodiment, the Pdcatalyst is Pd-G3, Pd₂(dba)₃, PdCl₂(MeCN)₂, Pd(OAc)₂, Pd(PPh₃)₄, orPdCl₂(PPh₃)₂. In one embodiment, the palladium catalyst is PdCl₂(MeCN)₂.In one embodiment, the palladium catalyst is Pd₂(dba)₃.

In one embodiment, the catalyst is a Ni catalyst. In one embodiment, theNi catalyst is (Ph₃P)₂NiCl₂.

In one embodiment, the catalyst is a Cu catalyst. In one embodiment, theCu catalyst is CuI.

In one embodiment, the ligand is a phosphine ligand or bisphosphineligand. In one embodiment, the ligand is XPhos, PCy₃, PCy₂Ph, P^(i)Pr₃,PCy₂ ^(t)Bu, CataCXium A, P(MeOC₆H₄)₃, PPh₂(C₆H₄CO₂H), PPh₂(C₆H₄SO₃H),SPhos, JohnPhos, DavePhos, MePhos, cBRIDP, Cy-vBRIDP, Cy-cBRIDP, ^(i)BuTriplecage, P^(t)Bu₂Cy, P^(t)Bu₃, CataCXium PICy, P^(t)Bu₂(PhNMe₂),PPh₃, dppp, dppe, dppb, BINAP, DPEPhos, dppf, dbpf, XantPhos, N-^(t)Bu₂Pazetine, dppm, dmpe, dippe, DIPAMP, Chiraphos, SPANphos, SEGPHOS,Me-DuPhos, or Josiphos. In one embodiment, the ligand is XPhos,CataCXium A, JohnPhos, DavePhos, MePhos, cBRIDP, CataCXium PICy, ordbpf. In one embodiment, wherein the ligand is XPhos(2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl). In oneembodiment, the ligand is cBRIDP.

In one embodiment, the molar ratio of the ligand to the catalyst is fromabout 5:1 to about 1:5. In one embodiment, the molar ratio of the ligandto the catalyst is from about 2:1 to about 1:2. In one embodiment, themolar ratio of the ligand to the catalyst is from about 2:1 to about1:1. In one embodiment, the ligand is a monodentate ligand and the molarratio of the ligand to the catalyst is about 2:1. In one embodiment, theligand is a monodentate ligand and the molar ratio of the ligand to thecatalyst is about 1:1. In one embodiment, the ligand is a bidentateligand and the molar ratio of the ligand to the catalyst is about 1:1.In one embodiment, the ligand is a bidentate ligand and the molar ratioof the ligand to the catalyst is about 1:2.

In one embodiment, the loading of the catalyst is from about 0.5% toabout 10%, from about 1% to about 10%, or from about 1% to about 5%. Inone embodiment, the loading of the catalyst is about 5%. In oneembodiment, the loading of the catalyst is about 4%. In one embodiment,the loading of the catalyst is about 3%. In one embodiment, the loadingof the catalyst is about 2%. In one embodiment, the loading of thecatalyst is about 1%.

In one embodiment, the loading of the ligand is from about 0.5% to about20%, from about 0.5% to about 15%, from about 0.5% to about 10%, fromabout 1% to about 10%, from about 1% to about 5%, or from about 1% toabout 3%. In one embodiment, the loading of the catalyst is about 10%.In one embodiment, the loading of the catalyst is about 5%. In oneembodiment, the loading of the catalyst is about 4%. In one embodiment,the loading of the catalyst is about 3%. In one embodiment, the loadingof the catalyst is about 2%. In one embodiment, the loading of thecatalyst is about 1%.

In one embodiment, the base is an inorganic base. In one embodiment, thebase is an alkali metal salt. In one embodiment, the base is an alkalineearth metal salt. In one embodiment, the base is Cs₂CO₃, K₂CO₃, orK₃PO₄. In one embodiment, the base is Cs₂CO₃. In one embodiment, thebase is K₂CO₃. In one embodiment, the base is K₃PO₄.

In one embodiment, the base is an organic base.

In one embodiment, the Pd catalyst is Pd₂(dba)₃, the ligand is Xphos,and the base is K₂CO₃. In one embodiment, the Pd catalyst is Pd₂(dba)₃,the ligand is Xphos, and the base is K₃PO₄. In one embodiment, the Pdcatalyst is PdCl₂(MeCN)₂, the ligand is Xphos, and the base is K₂CO₃. Inone embodiment, the Pd catalyst is Pd(OAc)₂, the ligand is Xphos, andthe base is K₂CO₃.

In one embodiment, the solvent is MeCN, ^(i)PrOAc, n-propyl acetate,2-MeTHF, EtCN, MEK, or toluene. In one embodiment, the solvent is MeCN.

In one embodiment, the coupling occurs in the presence of PdCl₂(MeCN)₂.In one embodiment, the coupling occurs further in the presence of XPhos.In one embodiment, the coupling occurs further in the presence of abase. In one embodiment, the base is Cs₂CO₃. In one embodiment, thecoupling occurs in the presence of a solvent. In one embodiment, thesolvent is an organic solvent. In one embodiment, the organic solvent isacetonitrile.

In one embodiment, the coupling occurs in the presence of Pd₂(dba)₃. Inone embodiment, the coupling occurs further in the presence of XPhos. Inone embodiment, the coupling occurs further in the presence of a base.In one embodiment, the base is K₃PO₄. In one embodiment, the couplingoccurs in the presence of a solvent. In one embodiment, the solvent isan organic solvent. In one embodiment, the organic solvent isacetonitrile.

In certain embodiments, provided herein is a process of preparing analkyne of Formula E:

comprising deprotecting a compound of Formula F:

In one embodiment, the deprotection occurs in the presence of a base anda solvent.

In one embodiment, the base is an inorganic base. In one embodiment, thebase is an alkali metal salt. In one embodiment, the base is KOH, NaOH,NaHCO₃, K₃PO₄, or K₂CO₃. In one embodiment, the base is an organic base.In one embodiment, the base is pyridine.

In one embodiment, the deprotection occurs in the presence of an acid.In one embodiment, the acid is HCl, AcOH, p-TsOH, or camphorsulfonicacid.

In one embodiment, the deprotection occurs in the presence of a fluoridesource. In one embodiment, the fluoride source is tetra-n-butylammoniumfluoride (TBAF). In one embodiment, the fluoride source is pyridine-HF.

In one embodiment, the deprotection occurs in the presence of aphase-transfer catalyst. In one embodiment, the phase-transfer catalystis tetrabutylammonium hydroxide.

In one embodiment, the solvent is an organic solvent. In one embodiment,the organic solvent is MeOH. In one embodiment, the solvent is a mixtureof water and a water immiscible solvent. In one embodiment, the waterimmiscible solvent is MTBE or DCM. In one embodiment, the deprotectionoccurs in the presence of a mixture of about 10 wt % KOH aqueoussolution and MTBE. In one embodiment, the solvent is acetonitrile.

In certain embodiments, provided herein is a process of preparing acompound of Formula F:

comprising coupling 4-iodo-1-methyl-1H-pyrazole withtrimethylsilylacetylene.

In one embodiment, the coupling occurs in the presence of a Cu catalyst,a Pd catalyst, and a base.

In one embodiment, the Cu catalyst is CuI.

In one embodiment, the Pd catalyst is Pd-G3, Pd₂(dba)₃, PdCl₂(MeCN)₂,Pd(OAc)₂, Pd(PPh₃)₄, or PdCl₂(PPh₃)₂. In one embodiment, the Pd catalystis PdCl₂(PPh₃)₂.

In one embodiment, the molar ratio of the Pd catalyst to the Cu catalystis from about 1:20 to about 10:1, from about 1:10 to about 5:1, fromabout 1:7.5 to about 1:1, from about 1:6 to about 1:2. In oneembodiment, the molar ratio of the Pd catalyst to the Cu catalyst isabout 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about1:4, about 1:3, about 1:2, about 1:1, or about 2:1. In one embodiment,the molar ratio of the Pd catalyst to the Cu catalyst is about 1:6. Inone embodiment, the molar ratio of the Pd catalyst to the Cu catalyst isabout 1:2.

In one embodiment, the loading of the Pd catalyst is from about 0.0005equivalent to about 0.1 equivalent, from about 0.001 equivalent to about0.05 equivalent, from about 0.002 equivalent to about 0.02 equivalent,or from about 0.003 equivalent to about 0.01 equivalent. In oneembodiment, the loading of the Pd catalyst is about 0.003 equivalent. Inone embodiment, the loading of the Pd catalyst is about 0.01 equivalent.

In one embodiment, the loading of the Cu catalyst is from about 0.001equivalent to about 0.2 equivalent, from about 0.005 equivalent to about0.1 equivalent, from about 0.01 equivalent to about 0.05 equivalent, orfrom about 0.0175 equivalent to about 0.02 equivalent. In oneembodiment, the loading of the Cu catalyst is about 0.0175 equivalent.In one embodiment, the loading of the Cu catalyst is about 0.02equivalent.

In one embodiment, the loading of the Pd catalyst is about 0.01equivalent and the loading of the Cu catalyst if about 0.02 equivalent.In one embodiment, the loading of the Pd catalyst is about 0.003equivalent and the loading of the Cu catalyst if about 0.0175equivalent.

In one embodiment, the base is DIPA, DIPEA, or N-methylmorpholine (NMM).In one embodiment, the base is DIPA. In one embodiment, the base isDIPEA. In one embodiment, the base is NMM. In one embodiment, the base(e.g., DIPA) is also used as the solvent.

In one embodiment, the coupling occurs in a solvent of DCM, toluene,2-methyl-tetrahydrofuran, or DIPA, or a mixture thereof. In oneembodiment, the coupling occurs in a solvent of DCM. In one embodiment,the coupling occurs in a solvent of toluene. In one embodiment, thecoupling occurs in a solvent of 2-methyl-tetrahydrofuran.

In one embodiment, the compound of Formula F is used in the preparationof the alkyne compound of Formula E without purification. In oneembodiment, the compound of Formula F is purified before being used inthe preparation of the alkyne compound of Formula E.

In one embodiment, the coupling produces less than about 5%, less thanabout 4%, less than about 3%, less than about 2%, less than about 1%,less than about 0.5%, less than about 0.3%, less than about 0.2%, orless than about 0.1% of a side-product of the formula

In certain embodiments, the carboxylic acid of Formula G is preparedaccording to the process as described in PCT publication Nos. WO2011/003065 and WO 2015/073267. An exemplary synthetic scheme is shownbelow. The overall average yield is from about 10% to about 25%.

In certain embodiments, provided herein is an alternative approach forthe preparation of the carboxylic acid of Formula G. An exemplarysynthetic scheme is shown below. This alternative approach results anoverall yield of about 40-45% and the product is obtained in an offwhite color.

In certain embodiments, the carboxylic acid of Formula G is prepared bya process comprising hydrolyzing a compound of Formula H:

In one embodiment, the hydrolysis occurs in the presence of a base. Inone embodiment, the base is LiOH, NaOH, or KOH. In one embodiment, thebase is LiOH.

In certain embodiments, the compound of Formula H is prepared by aprocess comprising reacting a compound of Formula J:

with 1,1,3,3-tetramethoxypropane.

In one embodiment, the reaction between the compound of Formula J and1,1,3,3-tetramethoxypropane occurs in a solvent of AcOH. In oneembodiment, the compound of Formula H is used in the preparation of thecompound of Formula G without further purification after the removal ofthe solvent of AcOH.

In one embodiment, the reaction between the compound of Formula J and1,1,3,3-tetramethoxypropane occurs in the presence of HCl.

In certain embodiment, provided herein is a process of preparing acompound of Formula J:

comprising cyclizing a compound of Formula K:

In one embodiment, the cyclization occurs by refluxing in a solvent of1-propanol for from about 2 days to about 4 days. In one embodiment, thecyclization occurs by refluxing in a solvent of 1-propanol for aboutdays.

In one embodiment, the cyclization occurs by refluxing in a solvent of1-butanol for from about 24 hours to about 48 hours. In one embodiment,the cyclization occurs by refluxing in a solvent of 1-butanol for about36 hours.

In one embodiment, the compound of Formula K is prepared by a processcomprising reacting a compound of Formula L:

with hydrazine or hydrazine hydrate.

In one embodiment, the reaction between the compound of Formula L andhydrazine or hydrazine hydrate occurs by refluxing in a solvent of1-propanol.

In one embodiment, the reaction between the compound of Formula L andhydrazine or hydrazine hydrate occurs by heating at from about 60° C. toabout 80° C. in a solvent of 1-butanol.

In one embodiment, the compound of Formula L is prepared by a processcomprising reacting a compound of Formula M:

with NH₃ or NH₄OH.

In one embodiment, the reaction between the compound of Formula M andNH₃ or NH₄OH occurs at room temperature.

In one embodiment, the preparations of the compound of Formula L, thecompound of Formula K, and the compound of Formula J occur in one-pot.

In one embodiment, the compound of Formula M is prepared by a processcomprising reacting ethyl 2-cyanoacetate with 2-chloroethylchloroformate.

In one embodiment, the reaction between 2-cyanoacetate and 2-chloroethylchloroformate occurs in the presence of a base. In one embodiment, thebase is an inorganic base. In one embodiment, the base is an alkalimetal salt. In one embodiment, the base is an alkaline earth metal salt.In one embodiment, the base is LiOH, NaOH, or KOH. In one embodiment,the base is NaOH.

In one embodiment, the reaction between 2-cyanoacetate and 2-chloroethylchloroformate occurs in a solvent of MeCN.

In one embodiment, the compound of Formula M is purified byrecrystalllization from MeOH. In one embodiment, the compound of FormulaM is purified by re-slurrying in 1-propanol. In one embodiment, thecompound of Formula M is purified by crystallization from 1-butanol.

5.4. Pharmaceutical Compositions

In some embodiments, provided herein are pharmaceutical compositionscomprising a solid form comprising a compound of Formula (I), or a salt,or solvate, or solvate of a salt thereof, or a mixture thereof, and abulking agent (filler or carrier), and optionally a disintegrant and alubricant. In some embodiments, provided herein are pharmaceuticalcompositions comprising a solid form provided herein, or a salt, orsolvate, or solvate of a salt thereof, or a mixture thereof, and apharmaceutically acceptable excipient, diluent, or carrier, includinginert solid diluents and fillers, sterile aqueous solution and variousorganic solvents, permeation enhancers, solubilizers and adjuvants. Inone embodiment, provided herein is a pharmaceutical compositioncomprising a solid form provided herein and a pharmaceutical acceptableexcipient thereof. In one embodiment, provided herein is apharmaceutical composition consisting essentially of a solid formprovided herein. In one embodiment, the solid form is present in saidcomposition in an amount of at least about 80% by weight. In oneembodiment, the solid form is present in said composition in an amountof at least about 90% by weight.

In one embodiment, the solid form in the pharmaceutical composition is apolymorphic or cocrystal form provided herein. In one embodiment, thesolid form is Form 1, Form 2, Form 3, Form 4, Form 5, Form 6, Form 7,Form 8, Form P1C3, Form P1C9, or Form P2C9 of Compound 1. In oneembodiment, the solid form is Form 1.

In one embodiment, the solid form in the pharmaceutical composition isan amorphous form of Compound 1. In one embodiment, the amorphous formof Compound 1 is prepared by a process provided herein. In oneembodiment, the amorphous form of Compound 1 is prepared by dissolvingForm 1 of Compound 1 one or more solvents to form a solution; andremoving the solvent of the solution to provide the amorphous form ofCompound 1. In one embodiment, the solvent is removed by spray drying.

In one embodiment, the pharmaceutical composition comprises one or moreexcipients selected from bulking agents (or fillers), disintegrants,lubricants, and capsule shell. In one embodiment, the bulking agent ismannitol or pre-gelatinized starch. In another embodiment, thedisintegrant is croscarmellose sodium. In another embodiment, thelubricant is magnesium stearate. In one embodiment, the capsule shell isHPMC capsule shell. In one embodiment, the pharmaceutical compositioncomprises one or more excipients selected from mannitol, pre-gelatinizedstarch, croscarmellose sodium, magnesium stearate, and HPMC capsuleshell.

In one embodiment, the amount of Compound 1 in the pharmaceuticalcomposition is about 1 mg to about 100 mg, about 1 mg to about 75 mg,about 1 mg to about 50 mg, about 1 mg to about 40 mg, about 5 mg toabout 50 mg, about 5 mg to about 30 mg, about 5 mg to about 10 mg, about5 mg, or about 30 mg. In one embodiment, the amount is about 5 mg, 10mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 75 mg, or 100 mg. In oneembodiment, the amount is about 5 mg or 30 mg. In one embodiment, theamount of Compound 1 in the pharmaceutical composition is about 1.5% toabout 25% w/w, about 1.5% to about 15% w/w, about 1.5% to about 10% w/w,about 1% to about 25% w/w, about 1% to about 15% w/w, or about 1% toabout 10% w/w. In one embodiment, the amount of Compound 1 in thepharmaceutical composition is about 1% to about 10% w/w. In oneembodiment, the amount of Compound 1 in the pharmaceutical compositionis about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5,9, 9.5, or 10% w/w. In one embodiment, the amount of Compound 1 in thepharmaceutical composition is about 1.9%, or about 9.4%. In oneembodiment, the amount of Compound 1 is about 1.92% or about 9.38%.

In one embodiment, the bulking agent (or filler) (e.g., starch andmannitol) in a pharmaceutical composition is about 80% to about 95% w/w,about 85% to about 95% w/w, or about 90% to about 95% w/w. In oneembodiment, the bulking agent (or filler) (e.g., starch and mannitol) ina pharmaceutical composition is about 80%, about 85%, about 90%, orabout 95% w/w. In one embodiment, the bulking agent (or filler) (e.g.,starch and mannitol) in a pharmaceutical composition is about 93% w/w,about 86% w/w, about 92.3% w/w, or about 85.1% w/w. In one embodiment,the bulking agent is about 93% w/w. In one embodiment, the bulking agentis about 85% w/w. In one embodiment, the bulking agent is starch,mannitol, or a mixture thereof. In one embodiment, the bulking agent isa mixture of starch and mannitol. In one embodiment, the weight ratio ofstarch to mannitol is from about 1:3 to about 3:1. In one embodiment,the bulking agent is an about 1:1 mixture of starch and mannitol. In oneembodiment, the starch is pre-gelatinized starch.

In one embodiment, the disintegrant (e.g., croscarmellose sodium) in apharmaceutical composition is about 1% to about 20% w/w, about 1% toabout 15% w/w, about 1% to about 10% w/w, about 2.5% to about 7.5% w/w,about 1% to about 5% w/w, or about 5% w/w. In one embodiment, thedisintegrant is about 1%, about 2%, about 3%, about 4%, about 5%, about6%, about 7%, about 8%, about 9%, or about 10% w/w. In one embodiment,the disintegrant is about 5% w/w.

In one embodiment, the lubricant (e.g., magnesium stearate) in apharmaceutical composition is about 0.1% to about 10% w/w, about 0.1% toabout 5% w/w, or about 0.1% to about 1% w/w. In one embodiment, thelubricant is about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%,about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1% w/w. In oneembodiment, the lubricant is about 0.5% w/w.

In one embodiment, provided herein is a process of preparing apharmaceutical composition provided herein comprising mixing a solidform comprising a compound of Formula (I) with a pharmaceuticallyacceptable excipient or carrier; wherein the solid form is Form 1, Form2, Form 3, Form 4, Form 5, Form 6, Form 7, Form 8, Form P1C3, Form P1C9,or Form P2C9. In one embodiment, provided herein is a pharmaceuticalcomposition prepared by the process above.

In one embodiment, provided herein is a pharmaceutical compositioncomprising an amorphous form of a compound of Formula (I), or a salt, orsolvate, or solvate of a salt thereof, or a mixture thereof, and abulking agent (filler or carrier), and optionally a disintegrant and alubricant. In one embodiment, provided herein is a pharmaceuticalcomposition comprising about 1% to about 10% w/w of an amorphous form ofa compound of Formula (I), or a salt, or solvate, or solvate of a saltthereof, or a mixture thereof, about 80% to about 95% w/w of a bulkingagent, about 2.5% to about 7.5% w/w of a disintegrant, and about 0.1% toabout 1% w/w of a lubricant.

In one embodiment, provided herein is a pharmaceutical compositioncomprising about 5 to 30 mg Compound 1 (e.g., amorphous),pre-gelatinized starch, and mannitol. In one embodiment, thepharmaceutical composition further comprises croscarmellose sodium andmagnesium stearate.

In one embodiment, the pharmaceutical composition is formulated asfollows: about 5 mg of Compound 1 (e.g., amorphous), about 120 mg ofpre-gelatinized starch, about 120 mg of mannitol, about 13 mg ofcroscarmellose sodium, and about 1.3 mg of magnesium stearate. Inembodiment, the pharmaceutical composition is formulated as a capsule.In one embodiment, the pharmaceutical composition is formulated asfollows: about 5 mg of Compound 1 (e.g., amorphous), about 120.35 mg ofpre-gelatinized starch, about 120.35 mg of mannitol, about 13.00 mg ofcroscarmellose sodium, and about 1.3 mg of magnesium stearate. Inembodiment, the pharmaceutical composition is formulated as a capsule.

In one embodiment, the pharmaceutical composition is formulated asfollows: about 30 mg of Compound 1 (e.g., amorphous), about 136 mg ofpre-gelatinized starch, about 136 mg of mannitol, about 16 mg ofcroscarmellose sodium, and about 1.6 mg of magnesium stearate. In oneembodiment, the pharmaceutical composition is formulated as follows:about 30 mg of Compound 1 (e.g., amorphous), about 136.20 mg ofpre-gelatinized starch, about 136.20 mg of mannitol, about 16.00 mg ofcroscarmellose sodium, and about 1.60 mg of magnesium stearate. Inembodiment, the pharmaceutical composition formulated as a capsule.

In some embodiments, a pharmaceutical composition described hereinincludes a second active agent such as an additional therapeutic agent,(e.g., a chemotherapeutic agent).

In some embodiments, provided herein is a pharmaceutical composition fororal administration (e.g., capsule) comprising: (a) about 5 mg ofamorphous Compound 1; (b) about 120.35 mg of pre-gelatinized starch; (c)about 120.35 mg of maanitol; (d) about 13 mg of croscarmellose sodium;and (e) about 1.3 mg of magnesium stearate.

In some embodiments, provided herein is a pharmaceutical composition fororal administration (e.g., capsule) comprising: (a) about 30 mg ofamorphous Compound 1; (b) about 136.2 mg of pre-gelatinized starch; (c)about 136.2 mg of maanitol; (d) about 16 mg of croscarmellose sodium;and (e) about 1.6 mg of magnesium stearate.

In one embodiment, the pharmaceutical composition is an oral dosageform. In one embodiment, the oral dosage form is a capsule. In anotherembodiment, the oral dosage form is a tablet. In one embodiment, thecapsule shell is Swedish orange or white.

5.4.1. Formulations

Pharmaceutical compositions can be specially formulated foradministration in solid or liquid form, including those adapted for thefollowing: oral administration, for example, drenches (aqueous ornon-aqueous solutions or suspensions), tablets (e.g., those targeted forbuccal, sublingual, and systemic absorption), capsules, boluses,powders, granules, pastes for application to the tongue, andintraduodenal routes; parenteral administration, including intravenous,intraarterial, subcutaneous, intramuscular, intravascular,intraperitoneal or infusion as, for example, a sterile solution orsuspension, or sustained-release formulation; topical application, forexample, as a cream, ointment, or a controlled-release patch or sprayapplied to the skin; intravaginally or intrarectally, for example, as apessary, cream, stent or foam; sublingually; ocularly; pulmonarily;local delivery by catheter or stent; intrathecally, or nasally.

Examples of suitable aqueous and nonaqueous carriers which can beemployed in pharmaceutical compositions include water, ethanol, polyols(such as glycerol, propylene glycol, polyethylene glycol, and the like),and suitable mixtures thereof, vegetable oils, such as olive oil, andinjectable organic esters, such as ethyl oleate. Proper fluidity can bemaintained, for example, by the use of coating materials, such aslecithin, by the maintenance of the required particle size in the caseof dispersions, and by the use of surfactants.

In one embodiment, suitable carriers which can be employed inpharmaceutical compositions include mannitol, pre-gelatinized starch,croscarmellose sodium, magnesium stearate, and HPMC capsule shell.

These compositions can also contain adjuvants such as preservatives,wetting agents, emulsifying agents, dispersing agents, lubricants,and/or antioxidants. Prevention of the action of microorganisms upon thecompounds described herein can be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It can also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form can be brought about by the inclusionof agents which delay absorption such as aluminum monostearate andgelatin.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound described herein and/or thechemotherapeutic with the carrier and, optionally, one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing into association a compound as provided herein withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Preparations for such pharmaceutical compositions are well-known in theart. See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, WilliamG, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill,2002; Pratt and Taylor, eds., Principles of Drug Action, Third Edition,Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and ClinicalPharmacology, Twelfth Edition, McGraw Hill, 2011; Goodman and Gilman,eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGrawHill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., LippincottWilliams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia,Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all ofwhich are incorporated by reference herein in their entirety. Exceptinsofar as any conventional excipient medium is incompatible with thecompounds provided herein, such as by producing any undesirablebiological effect or otherwise interacting in a deleterious manner withany other component(s) of the pharmaceutically acceptable composition,the excipient's use is contemplated to be within the scope of thisdisclosure.

In some embodiments, the concentration of one or more of the compoundsprovided in the disclosed pharmaceutical compositions is equal to orless than about 100%, about 90%, about 80%, about 70%, about 60%, about50%, about 40%, about 30%, about 20%, about 19%, about 18%, about 17%,about 16%, about 15%, about 14%, about 13%, about 12%, about 11%, about10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about3%, about 2%, about 1%, about 0.5%, about 0.4%, about 0.3%, about 0.2%,about 0.1%, about 0.09%, about 0.08%, about 0.07%, about 0.06%, about0.05%, about 0.04%, about 0.03%, about 0.02%, about 0.01%, about 0.009%,about 0.008%, about 0.007%, about 0.006%, about 0.005%, about 0.004%,about 0.003%, about 0.002%, about 0.001%, about 0.0009%, about 0.0008%,about 0.0007%, about 0.0006%, about 0.0005%, about 0.0004%, about0.0003%, about 0.0002%, or about 0.0001% w/w, w/v or v/v.

In some embodiments, the concentration of one or more of the compoundsas provided herein is greater than about 90%, about 80%, about 70%,about 60%, about 50%, about 40%, about 30%, about 20%, about 19.75%,about 19.50%, about 19.25%, about 19%, about 18.75%, about 18.50%, about18.25%, about 18%, about 17.75%, about 17.50%, about 17.25%, about 17%,about 16.75%, about 16.50%, about 16.25%, about 16%, about 15.75%, about15.50%, about 15.25%, about 15%, about 14.75%, about 14.50%, about14.25%, about 14%, about 13.75%, about 13.50%, about 13.25%, about 13%,about 12.75%, about 12.50%, about 12.25%, about 12%, about 11.75%, about11.50%, about 11.25%, about 11%, about 10.75%, about 10.50%, about10.25%, about 10%, about 9.75%, about 9.50%, about 9.25%, about 9%,about 8.75%, about 8.50%, about 8.25%, about 8%, about 7.75%, about7.50%, about 7.25%, about 7%, about 6.75%, about 6.50%, about 6.25%,about 6%, about 5.75%, about 5.50%, about 5.25%, about 5%, about 4.75%,about 4.50%, about 4.25%, about 4%, about 3.75%, about 3.50%, about3.25%, about 3%, about 2.75%, about 2.50%, about 2.25%, about 2%, about1.75%, about 1.50%, about 1.25%, about 1%, about 0.5%, about 0.4%, about0.3%, about 0.2%, about 0.1%, about 0.09%, about 0.08%, about 0.07%,about 0.06%, about 0.05%, about 0.04%, about 0.03%, about 0.02%, about0.01%, about 0.009%, about 0.008%, about 0.007%, about 0.006%, about0.005%, about 0.004%, about 0.003%, about 0.002%, about 0.001%, about0.0009%, about 0.0008%, about 0.0007%, about 0.0006%, about 0.0005%,about 0.0004%, about 0.0003%, about 0.0002%, or about 0.0001% w/w, w/v,or v/v.

In some embodiments, the concentration of one or more of the compoundsas provided herein is in the range from approximately 0.0001% toapproximately 50%, approximately 0.001% to approximately 40%,approximately 0.01% to approximately 30%, approximately 0.02% toapproximately 29%, approximately 0.03% to approximately 28%,approximately 0.04% to approximately 27%, approximately 0.05% toapproximately 26%, approximately 0.06% to approximately 25%,approximately 0.07% to approximately 24%, approximately 0.08% toapproximately 23%, approximately 0.09% to approximately 22%,approximately 0.1% to approximately 21%, approximately 0.2% toapproximately 20%, approximately 0.3% to approximately 19%,approximately 0.4% to approximately 18%, approximately 0.5% toapproximately 17%, approximately 0.6% to approximately 16%,approximately 0.7% to approximately 15%, approximately 0.8% toapproximately 14%, approximately 0.9% to approximately 12%, orapproximately 1% to approximately 10% w/w, w/v, or v/v.

In some embodiments, the concentration of one or more of the compoundsas provided herein is in the range from approximately 0.001% toapproximately 10%, approximately 0.01% to approximately 5%,approximately 0.02% to approximately 4.5%, approximately 0.03% toapproximately 4%, approximately 0.04% to approximately 3.5%,approximately 0.05% to approximately 3%, approximately 0.06% toapproximately 2.5%, approximately 0.07% to approximately 2%,approximately 0.08% to approximately 1.5%, approximately 0.09% toapproximately 1%, or approximately 0.1% to approximately 0.9% w/w, w/v,or v/v.

In some embodiments, the amount of one or more of the compounds asprovided herein is equal to or less than about 10 g, about 9.5 g, about9.0 g, about 8.5 g, about 8.0 g, about 7.5 g, about 7.0 g, about 6.5 g,about 6.0 g, about 5.5 g, about 5.0 g, about 4.5 g, about 4.0 g, about3.5 g, about 3.0 g, about 2.5 g, about 2.0 g, about 1.5 g, about 1.0 g,about 0.95 g, about 0.9 g, about 0.85 g, about 0.8 g, about 0.75 g,about 0.7 g, about 0.65 g, about 0.6 g, about 0.55 g, about 0.5 g, about0.45 g, about 0.4 g, about 0.35 g, about 0.3 g, about 0.25 g, about 0.2g, about 0.15 g, about 0.1 g, about 0.09 g, about 0.08 g, about 0.07 g,about 0.06 g, about 0.05 g, about 0.04 g, about 0.03 g, about 0.02 g,about 0.01 g, about 0.009 g, about 0.008 g, about 0.007 g, about 0.006g, about 0.005 g, about 0.004 g, about 0.003 g, about 0.002 g, about0.001 g, about 0.0009 g, about 0.0008 g, about 0.0007 g, about 0.0006 g,about 0.0005 g, about 0.0004 g, about 0.0003 g, about 0.0002 g, or about0.0001 g. In some embodiments, the amount of one or more of thecompounds provided herein in the pharmaceutical compositions providedherein is about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3mg, about 3.1 mg, about 3.2 mg, about 3.3 mg, about 3.4 mg, about 3.5mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9mg, or about 10 mg.

In some embodiments, the amount of one or more of the compounds asprovided herein is more than about 0.0001 g, about 0.0002 g, about0.0003 g, about 0.0004 g, about 0.0005 g, about 0.0006 g, about 0.0007g, about 0.0008 g, about 0.0009 g, about 0.001 g, about 0.0015 g, about0.002 g, about 0.0025 g, about 0.003 g, about 0.0035 g, about 0.004 g,about 0.0045 g, about 0.005 g, about 0.0055 g, about 0.006 g, about0.0065 g, about 0.007 g, about 0.0075 g, about 0.008 g, about 0.0085 g,about 0.009 g, about 0.0095 g, about 0.01 g, about 0.015 g, about 0.02g, about 0.025 g, about 0.03 g, about 0.035 g, about 0.04 g, about 0.045g, about 0.05 g, about 0.055 g, about 0.06 g, about 0.065 g, about 0.07g, about 0.075 g, about 0.08 g, about 0.085 g, about 0.09 g, about 0.095g, about 0.1 g, about 0.15 g, about 0.2 g, about 0.25 g, about 0.3 g,about 0.35 g, about 0.4 g, about 0.45 g, about 0.5 g, about 0.55 g,about 0.6 g, about 0.65 g, about 0.7 g, about 0.75 g, about 0.8 g, about0.85 g, about 0.9 g, about 0.95 g, about 1 g, about 1.5 g, about 2 g,about 2.5 g, about 3 g, about 3.5 g, about 4 g, about 4.5 g, about 5 g,about 5.5 g, about 6 g, about 6.5 g, about 7 g, about 7.5 g, about 8 g,about 8.5 g, about 9 g, about 9.5 g, or about 10 g.

In some embodiments, the amount of one or more of the compounds asprovided herein is in the range of about 0.0001 to about 10 g, about0.0005 to about 9 g, about 0.001 to about 8 g, about 0.005 to about 7 g,about 0.01 to about 6 g, about 0.05 to about 5 g, about 0.1 to about 4g, about 0.5 to about 4 g, or about 1 to about 3 g.

5.4.1.1 Formulations for Oral Administration

In some embodiments, provided herein are pharmaceutical compositions fororal administration containing a compound as provided herein, and apharmaceutical excipient suitable for oral administration. In someembodiments, provided herein are pharmaceutical compositions for oraladministration containing: (i) an effective amount of a disclosedcompound; optionally (ii) an effective amount of one or more secondagents; and (iii) one or more pharmaceutical excipients suitable fororal administration. In some embodiments, the pharmaceutical compositionfurther contains: (iv) an effective amount of a third agent.

In some embodiments, the pharmaceutical composition can be a liquidpharmaceutical composition suitable for oral consumption. Pharmaceuticalcompositions suitable for oral administration can be presented asdiscrete dosage forms, such as capsules, cachets, or tablets, or liquidsor aerosol sprays each containing a predetermined amount of an activeingredient as a powder or in granules, a solution, or a suspension in anaqueous or non-aqueous liquid, an oil-in-water emulsion, or awater-in-oil liquid emulsion. Such dosage forms can be prepared by anyof the methods of pharmacy, but all methods include the step of bringingthe active ingredient into association with the carrier, whichconstitutes one or more ingredients. In general, the pharmaceuticalcompositions are prepared by uniformly and intimately admixing theactive ingredient with liquid carriers or finely divided solid carriersor both, and then, if necessary, shaping the product into the desiredpresentation. For example, a tablet can be prepared by compression ormolding, optionally with one or more accessory ingredients. Compressedtablets can be prepared by compressing in a suitable machine the activeingredient in a free-flowing form such as powder or granules, optionallymixed with an excipient such as, but not limited to, a binder, alubricant, an inert diluent, and/or a surface active or dispersingagent. Molded tablets can be made by molding in a suitable machine amixture of the powdered compound moistened with an inert liquid diluent.

The present disclosure further encompasses anhydrous pharmaceuticalcompositions and dosage forms comprising an active ingredient, sincewater can facilitate the degradation of some compounds. For example,water can be added (e.g., about 5%) in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. Anhydrous pharmaceutical compositions and dosage forms can beprepared using anhydrous or low moisture containing ingredients and lowmoisture or low humidity conditions. For example, pharmaceuticalcompositions and dosage forms which contain lactose can be madeanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected. An anhydrouspharmaceutical composition can be prepared and stored such that itsanhydrous nature is maintained. Accordingly, anhydrous pharmaceuticalcompositions can be packaged using materials known to prevent exposureto water such that they can be included in suitable formulary kits.Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastic or the like, unit dose containers,blister packs, and strip packs.

An active ingredient can be combined in an intimate admixture with apharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier can take a wide variety of formsdepending on the form of preparation desired for administration. Inpreparing the pharmaceutical compositions for an oral dosage form, anyof the usual pharmaceutical media can be employed as carriers, such as,for example, water, glycols, oils, alcohols, flavoring agents,preservatives, coloring agents, and the like in the case of oral liquidpreparations (such as suspensions, solutions, and elixirs) or aerosols;or carriers such as starches, sugars, micro-crystalline cellulose,diluents, granulating agents, lubricants, binders, and disintegratingagents can be used in the case of oral solid preparations, in someembodiments without employing the use of lactose. For example, suitablecarriers include powders, capsules, and tablets, with the solid oralpreparations. In some embodiments, tablets can be coated by standardaqueous or nonaqueous techniques.

Binders suitable for use in pharmaceutical compositions and dosage formsinclude, but are not limited to, corn starch, potato starch, or otherstarches, gelatin, natural and synthetic gums such as acacia, sodiumalginate, alginic acid, other alginates, powdered tragacanth, guar gum,cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate,carboxymethyl cellulose calcium, sodium carboxymethyl cellulose),polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch,hydroxypropyl methyl cellulose, microcrystalline cellulose, and mixturesthereof.

Examples of suitable fillers for use in the pharmaceutical compositionsand dosage forms provided herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants can be used in the pharmaceutical compositions as providedherein to provide tablets that disintegrate when exposed to an aqueousenvironment. Too much of a disintegrant can produce tablets which candisintegrate in the bottle. Too little can be insufficient fordisintegration to occur and can thus alter the rate and extent ofrelease of the active ingredient(s) from the dosage form. Thus, asufficient amount of disintegrant that is neither too little nor toomuch to detrimentally alter the release of the active ingredient(s) canbe used to form the dosage forms of the compounds provided herein. Theamount of disintegrant used can vary based upon the type of formulationand mode of administration, and can be readily discernible to those ofordinary skill in the art. About 0.5 to about 15 weight percent ofdisintegrant, or about 1 to about 5 weight percent of disintegrant, canbe used in the pharmaceutical composition. Disintegrants that can beused to form pharmaceutical compositions and dosage forms include, butare not limited to, agar-agar, alginic acid, calcium carbonate,microcrystalline cellulose, croscarmellose sodium, crospovidone,polacrilin potassium, sodium starch glycolate, potato or tapioca starch,other starches, pre-gelatinized starch, other starches, clays, otheralgins, other celluloses, gums or mixtures thereof.

Lubricants which can be used to form pharmaceutical compositions anddosage forms include, but are not limited to, calcium stearate,magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol,mannitol, polyethylene glycol, other glycols, stearic acid, sodiumlauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil,cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethylaureate, agar, ormixtures thereof. Additional lubricants include, for example, a syloidsilica gel, a coagulated aerosol of synthetic silica, or mixturesthereof. A lubricant can optionally be added, in an amount of less thanabout 1 weight percent of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oraladministration, the active ingredient therein can be combined withvarious sweetening or flavoring agents, coloring matter or dyes and, forexample, emulsifying and/or suspending agents, together with suchdiluents as water, ethanol, propylene glycol, glycerin and variouscombinations thereof.

The tablets can be uncoated or coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin or olive oil.

Surfactant which can be used to form pharmaceutical compositions anddosage forms include, but are not limited to, hydrophilic surfactants,lipophilic surfactants, and mixtures thereof. That is, a mixture ofhydrophilic surfactants can be employed, a mixture of lipophilicsurfactants can be employed, or a mixture of at least one hydrophilicsurfactant and at least one lipophilic surfactant can be employed.

A suitable hydrophilic surfactant can generally have an HLB value of atleast about 10, while suitable lipophilic surfactants can generally havean HLB value of or less than about 10. An empirical parameter used tocharacterize the relative hydrophilicity and hydrophobicity of non-ionicamphiphilic compounds is the hydrophilic-lipophilic balance (“HLB”value). Surfactants with lower HLB values are more lipophilic orhydrophobic, and have greater solubility in oils, while surfactants withhigher HLB values are more hydrophilic, and have greater solubility inaqueous solutions. Hydrophilic surfactants are generally considered tobe those compounds having an HLB value greater than about 10, as well asanionic, cationic, or zwitterionic compounds for which the HLB scale isnot generally applicable. Similarly, lipophilic (i.e., hydrophobic)surfactants are compounds having an HLB value equal to or less thanabout 10. However, HLB value of a surfactant is merely a rough guidegenerally used to enable formulation of industrial, pharmaceutical andcosmetic emulsions.

Hydrophilic surfactants can be either ionic or non-ionic. Suitable ionicsurfactants include, but are not limited to, alkylammonium salts;fusidic acid salts; fatty acid derivatives of amino acids,oligopeptides, and polypeptides; glyceride derivatives of amino acids,oligopeptides, and polypeptides; lecithins and hydrogenated lecithins;lysolecithins and hydrogenated lysolecithins; phospholipids andderivatives thereof lysophospholipids and derivatives thereof; carnitinefatty acid ester salts; salts of alkylsulfates; fatty acid salts; sodiumdocusate; acylactylates; mono- and di-acetylated tartaric acid esters ofmono- and di-glycerides; succinylated mono- and di-glycerides; citricacid esters of mono- and di-glycerides; and mixtures thereof.

Within the aforementioned group, ionic surfactants include, by way ofexample: lecithins, lysolecithin, phospholipids, lysophospholipids andderivatives thereof carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; and mixtures thereof.

Ionic surfactants can be the ionized forms of lecithin, lysolecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol,phosphatidic acid, phosphatidylserine, lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine, lactylic esters of fatty acids,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants can include, but are not limited to,alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esterssuch as polyethylene glycol fatty acids monoesters and polyethyleneglycol fatty acids diesters; polyethylene glycol glycerol fatty acidesters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fattyacid esters such as polyethylene glycol sorbitan fatty acid esters;hydrophilic transesterification products of a polyol with at least onemember of glycerides, vegetable oils, hydrogenated vegetable oils, fattyacids, and sterols; polyoxyethylene sterols, derivatives, and analoguesthereof; polyoxyethylated vitamins and derivatives thereof;polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof;polyethylene glycol sorbitan fatty acid esters and hydrophilictransesterification products of a polyol with at least one member oftriglycerides, vegetable oils, and hydrogenated vegetable oils. Thepolyol can be glycerol, ethylene glycol, polyethylene glycol, sorbitol,propylene glycol, pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate,PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate,PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryllaurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenatedcastor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate,sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG 15-100 octylphenol series, and poloxamers.

Suitable lipophilic surfactants include, by way of example only: fattyalcohols; glycerol fatty acid esters; acetylated glycerol fatty acidesters; lower alcohol fatty acids esters; propylene glycol fatty acidesters; sorbitan fatty acid esters; polyethylene glycol sorbitan fattyacid esters; sterols and sterol derivatives; polyoxyethylated sterolsand sterol derivatives; polyethylene glycol alkyl ethers; sugar esters;sugar ethers; lactic acid derivatives of mono- and di-glycerides;hydrophobic transesterification products of a polyol with at least onemember of glycerides, vegetable oils, hydrogenated vegetable oils, fattyacids and sterols; oil-soluble vitamins/vitamin derivatives; andmixtures thereof. Within this group, non-limiting examples of lipophilicsurfactants include glycerol fatty acid esters, propylene glycol fattyacid esters, and mixtures thereof, or are hydrophobictransesterification products of a polyol with at least one member ofvegetable oils, hydrogenated vegetable oils, and triglycerides.

In one embodiment, the pharmaceutical composition can include asolubilizer to ensure good solubilization and/or dissolution of acompound as provided herein and to minimize precipitation of thecompound. This can be especially important for pharmaceuticalcompositions for non-oral use, e.g., pharmaceutical compositions forinjection. A solubilizer can also be added to increase the solubility ofthe hydrophilic drug and/or other components, such as surfactants, or tomaintain the pharmaceutical composition as a stable or homogeneoussolution or dispersion.

Examples of suitable solubilizers include, but are not limited to, thefollowing: alcohols and polyols, such as ethanol, isopropyl alcohol,butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediolsand isomers thereof, glycerol, pentaerythritol, sorbitol, mannitol,transcutol, dimethyl isosorbide, polyethylene glycol, polypropyleneglycol, polyvinylalcohol, hydroxypropyl methylcellulose and othercellulose derivatives, cyclodextrins and cyclodextrin derivatives;ethers of polyethylene glycols having an average molecular weight ofabout 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether(glycofurol) or methoxy PEG; amides and other nitrogen-containingcompounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam,N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone,N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esterssuch as ethyl propionate, tributylcitrate, acetyl triethylcitrate,acetyl tributyl citrate, triethylcitrate, ethyl oleate, ethyl caprylate,ethyl butyrate, triacetin, propylene glycol monoacetate, propyleneglycol diacetate, ε-caprolactone and isomers thereof, δ-valerolactoneand isomers thereof, β-butyrolactone and isomers thereof; and othersolubilizers known in the art, such as dimethyl acetamide, dimethylisosorbide, N-methyl pyrrolidones, monooctanoin, diethylene glycolmonoethyl ether, and water.

Mixtures of solubilizers can also be used. Examples include, but notlimited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol,transcutol, propylene glycol, and dimethyl isosorbide. In someembodiments, solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included is not particularlylimited. The amount of a given solubilizer can be limited to abioacceptable amount, which can be readily determined by one of skill inthe art. In some circumstances, it can be advantageous to includeamounts of solubilizers far in excess of bioacceptable amounts, forexample to maximize the concentration of the drug, with excesssolubilizer removed prior to providing the pharmaceutical composition toa subject using conventional techniques, such as distillation orevaporation. Thus, if present, the solubilizer can be in a weight ratioof about 10%, 25%, 50%, 100%, or up to about 200% by weight, based onthe combined weight of the drug, and other excipients. If desired, verysmall amounts of solubilizer can also be used, such as about 5%, 2%, 1%or even less. Typically, the solubilizer can be present in an amount ofabout 1% to about 100%, more typically about 5% to about 25% by weight.

The pharmaceutical composition can further include one or morepharmaceutically acceptable additives and excipients. Such additives andexcipients include, without limitation, detackifiers, anti-foamingagents, buffering agents, polymers, antioxidants, preservatives,chelating agents, viscomodulators, tonicifiers, flavorants, colorants,oils, odorants, opacifiers, suspending agents, binders, fillers,plasticizers, lubricants, and mixtures thereof.

Exemplary preservatives can include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives. Exemplaryantioxidants include, but are not limited to, alpha tocopherol, ascorbicacid, acorbyl palmitate, butylated hydroxyanisole, butylatedhydroxytoluene, monothioglycerol, potassium metabisulfite, propionicacid, propyl gallate, sodium ascorbate, sodium bisulfite, sodiummetabisulfite, and sodium sulfite. Exemplary chelating agents includeethylenediaminetetraacetic acid (EDTA), citric acid monohydrate,disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malicacid, phosphoric acid, sodium edetate, tartaric acid, and trisodiumedetate. Exemplary antimicrobial preservatives include, but are notlimited to, benzalkonium chloride, benzethonium chloride, benzylalcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine,chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol,glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethylalcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.Exemplary antifungal preservatives include, but are not limited to,butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoicacid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodiumbenzoate, sodium propionate, and sorbic acid. Exemplary alcoholpreservatives include, but are not limited to, ethanol, polyethyleneglycol, phenol, phenolic compounds, bisphenol, chlorobutanol,hydroxybenzoate, and phenylethyl alcohol. Exemplary acidic preservativesinclude, but are not limited to, vitamin A, vitamin C, vitamin E,beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbicacid, sorbic acid, and phytic acid. Other preservatives include, but arenot limited to, tocopherol, tocopherol acetate, deteroxime mesylate,cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened(BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ethersulfate (SLES), sodium bisulfite, sodium metabisulfite, potassiumsulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben,Germall 115, Germaben II, Neolone, Kathon, and Euxyl. In certainembodiments, the preservative is an anti-oxidant. In other embodiments,the preservative is a chelating agent.

Exemplary oils include, but are not limited to, almond, apricot kernel,avocado, babassu, bergamot, black current seed, borage, cade, camomile,canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, codliver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose,fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop,isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon,litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink,nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel,peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary,safflower, sandalwood, sasquana, savory, sea buckthorn, sesame, sheabutter, silicone, soybean, sunflower, tea tree, thistle, tsubaki,vetiver, walnut, and wheat germ oils. Exemplary oils also include, butare not limited to, butyl stearate, caprylic triglyceride, caprictriglyceride, cyclomethicone, diethyl sebacate, dimethicone 360,isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol,silicone oil, and combinations thereof.

In addition, an acid or a base can be incorporated into thepharmaceutical composition to facilitate processing, to enhancestability, or for other reasons. Examples of pharmaceutically acceptablebases include amino acids, amino acid esters, ammonium hydroxide,potassium hydroxide, sodium hydroxide, sodium hydrogen carbonate,aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesiumaluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite,magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine,ethylenediamine, triethanolamine, triethylamine, triisopropylalcoholamine, trimethylamine, tris(hydroxymethyl)-aminomethane (TRIS)and the like. Also suitable are bases that are salts of apharmaceutically acceptable acid, such as acetic acid, acrylic acid,adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbicacid, benzoic acid, boric acid, butyric acid, carbonic acid, citricacid, fatty acids, formic acid, fumaric acid, gluconic acid,hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid,oxalic acid, para-bromophenylsulfonic acid, propionic acid,p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid,tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid,uric acid, and the like. Salts of polyprotic acids, such as sodiumphosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphatecan also be used. When the base is a salt, the cation can be anyconvenient and pharmaceutically acceptable cation, such as ammonium,alkali metals, alkaline earth metals, and the like. Examples caninclude, but not limited to, sodium, potassium, lithium, magnesium,calcium and ammonium.

Suitable acids are pharmaceutically acceptable organic or inorganicacids. Examples of suitable inorganic acids include hydrochloric acid,hydrobromic acid, hydriodic acid, sulfuric acid, nitric acid, boricacid, phosphoric acid, and the like. Examples of suitable organic acidsinclude acetic acid, acrylic acid, adipic acid, alginic acid,alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boricacid, butyric acid, carbonic acid, citric acid, fatty acids, formicacid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbicacid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid,para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid,salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid,thioglycolic acid, toluenesulfonic acid, uric acid and the like.

5.4.1.2 Formulations for Parenteral Administration

In some embodiments, provided herein are pharmaceutical compositions forparenteral administration containing a compound as provided herein, anda pharmaceutical excipient suitable for parenteral administration. Insome embodiments, provided herein are pharmaceutical compositions forparenteral administration containing: (i) an effective amount of adisclosed compound; optionally (ii) an effective amount of one or moresecond agents; and (iii) one or more pharmaceutical excipients suitablefor parenteral administration. In some embodiments, the pharmaceuticalcomposition further contains: (iv) an effective amount of a third agent.

The forms in which the disclosed pharmaceutical compositions can beincorporated for administration by injection include aqueous or oilsuspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, orpeanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueoussolution, and similar pharmaceutical vehicles.

Aqueous solutions in saline are also conventionally used for injection.Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and thelike (and suitable mixtures thereof), cyclodextrin derivatives, andvegetable oils can also be employed.

Aqueous solutions in saline are also conventionally used for injection.Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, and thelike (and suitable mixtures thereof), cyclodextrin derivatives, andvegetable oils can also be employed. The proper fluidity can bemaintained, for example, by the use of a coating, such as lecithin, forthe maintenance of the required particle size in the case of dispersionand by the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating a compound asprovided herein in the required amount in the appropriate solvent withvarious other ingredients as enumerated above, as appropriate, followedby filtered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the appropriateother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, certainmethods of preparation are vacuum-drying and freeze-drying techniqueswhich yield a powder of the active ingredient plus any additionalingredient from a previously sterile-filtered solution thereof.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use. Injectable compositions can contain from about 0.1to about 5% w/w of a compound as disclosed herein.

5.4.1.3 Formulations for Topical Administration

In some embodiments, provided herein are pharmaceutical compositions fortopical (e.g., transdermal) administration containing a compound asprovided herein, and a pharmaceutical excipient suitable for topicaladministration. In some embodiments, provided herein are pharmaceuticalcompositions for topical administration containing: (i) an effectiveamount of a disclosed compound; optionally (ii) an effective amount ofone or more second agents; and (iii) one or more pharmaceuticalexcipients suitable for topical administration. In some embodiments, thepharmaceutical composition further contains: (iv) an effective amount ofa third agent.

Pharmaceutical compositions provided herein can be formulated intopreparations in solid, semi-solid, or liquid forms suitable for local ortopical administration, such as gels, water soluble jellies, creams,lotions, suspensions, foams, powders, slurries, ointments, solutions,oils, pastes, suppositories, sprays, emulsions, saline solutions,dimethylsulfoxide (DMSO)-based solutions. In general, carriers withhigher densities are capable of providing an area with a prolongedexposure to the active ingredients. In contrast, a solution formulationcan provide more immediate exposure of the active ingredient to thechosen area.

The pharmaceutical compositions also can comprise suitable solid or gelphase carriers or excipients, which are compounds that allow increasedpenetration of, or assist in the delivery of, therapeutic moleculesacross the stratum corneum permeability barrier of the skin. There aremany of these penetration-enhancing molecules known to those trained inthe art of topical formulation. Examples of such carriers and excipientsinclude, but are not limited to, humectants (e.g., urea), glycols (e.g.,propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleicacid), surfactants (e.g., isopropyl myristate and sodium laurylsulfate), pyrrolidones, glycerol monolaurate, sulfoxides, terpenes(e.g., menthol), amines, amides, alkanes, alkanols, water, calciumcarbonate, calcium phosphate, various sugars, starches, cellulosederivatives, gelatin, and polymers such as polyethylene glycols.

Another exemplary formulation for use in the disclosed methods employstransdermal delivery devices (“patches”). Such transdermal patches canbe used to provide continuous or discontinuous infusion of a compound asprovided herein in controlled amounts, either with or without anotheragent.

The construction and use of transdermal patches for the delivery ofpharmaceutical agents is well known in the art. See, e.g., U.S. Pat.Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches can be constructedfor continuous, pulsatile, or on demand delivery of pharmaceuticalagents.

Suitable devices for use in delivering intradermal pharmaceuticallyacceptable compositions described herein include short needle devicessuch as those described in U.S. Pat. Nos. 4,886,499; 5,190,521;5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662.Intradermal compositions can be administered by devices which limit theeffective penetration length of a needle into the skin, such as thosedescribed in PCT publication WO 99/34850 and functional equivalentsthereof. Jet injection devices which deliver liquid vaccines to thedermis via a liquid jet injector and/or via a needle which pierces thestratum corneum and produces a jet which reaches the dermis aresuitable. Jet injection devices are described, for example, in U.S. Pat.Nos. 5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189;5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335;5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880;4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballisticpowder/particle delivery devices which use compressed gas to acceleratevaccine in powder form through the outer layers of the skin to thedermis are suitable. Alternatively or additionally, conventionalsyringes can be used in the classical mantoux method of intradermaladministration.

Topically-administrable formulations can, for example, comprise fromabout 1% to about 10% (w/w) of a compound provided herein relative tothe total weight of the formulation, although the concentration of thecompound provided herein in the formulation can be as high as thesolubility limit of the compound in the solvent. In some embodiments,topically-administrable formulations can, for example, comprise fromabout 1% to about 9% (w/w) of a compound provided herein, such as fromabout 1% to about 8% (w/w), further such as from about 1% to about 7%(w/w), further such as from about 1% to about 6% (w/w), further such asfrom about 1% to about 5% (w/w), further such as from about 1% to about4% (w/w), further such as from about 1% to about 3% (w/w), and furthersuch as from about 1% to about 2% (w/w) of a compound provided herein.Formulations for topical administration can further comprise one or moreof the additional pharmaceutically acceptable excipients describedherein.

5.4.1.4 Formulations for Inhalation Administration

In some embodiments, provided herein are pharmaceutical compositions forinhalation administration containing a compound as provided herein, anda pharmaceutical excipient suitable for topical administration. In someembodiments, provided herein are pharmaceutical compositions forinhalation administration containing: (i) an effective amount of adisclosed compound; optionally (ii) an effective amount of one or moresecond agents; and (iii) one or more pharmaceutical excipients suitablefor inhalation administration. In some embodiments, the pharmaceuticalcomposition further contains: (iv) an effective amount of a third agent.

Pharmaceutical compositions for inhalation or insufflation includesolutions and suspensions in pharmaceutically acceptable, aqueous ororganic solvents, or mixtures thereof, and powders. The liquid or solidpharmaceutical compositions can contain suitable pharmaceuticallyacceptable excipients as described herein. In some embodiments, thepharmaceutical compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Pharmaceuticalcompositions in pharmaceutically acceptable solvents can be nebulized byuse of inert gases. Nebulized solutions can be inhaled directly from thenebulizing device or the nebulizing device can be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder pharmaceutical compositions can beadministered, e.g., orally or nasally, from devices that deliver theformulation in an appropriate manner.

5.4.1.5 Formulations for Ocular Administration

In some embodiments, the disclosure provides a pharmaceuticalcomposition for treating ophthalmic disorders. The pharmaceuticalcomposition can contain an effective amount of a compound as providedherein and a pharmaceutical excipient suitable for ocularadministration. Pharmaceutical compositions suitable for ocularadministration can be presented as discrete dosage forms, such as dropsor sprays each containing a predetermined amount of an active ingredienta solution, or a suspension in an aqueous or non-aqueous liquid, anoil-in-water emulsion, or a water-in-oil liquid emulsion. Otheradministration forms include intraocular injection, intravitrealinjection, topically, or through the use of a drug eluting device,microcapsule, implant, or microfluidic device. In some cases, thecompounds as provided herein are administered with a carrier orexcipient that increases the intraocular penetrance of the compound suchas an oil and water emulsion with colloid particles having an oily coresurrounded by an interfacial film. It is contemplated that all localroutes to the eye can be used including topical, subconjunctival,periocular, retrobulbar, subtenon, intracameral, intravitreal,intraocular, subretinal, juxtascleral and suprachoroidal administration.Systemic or parenteral administration can be feasible including, but notlimited to intravenous, subcutaneous, and oral delivery. An exemplarymethod of administration will be intravitreal or subtenon injection ofsolutions or suspensions, or intravitreal or subtenon placement ofbioerodible or non-bioerodible devices, or by topical ocularadministration of solutions or suspensions, or posterior juxtascleraladministration of a gel or cream formulation.

Eye drops can be prepared by dissolving the active ingredient in asterile aqueous solution such as physiological saline, bufferingsolution, etc., or by combining powder compositions to be dissolvedbefore use. Other vehicles can be chosen, as is known in the art,including, but not limited to: balance salt solution, saline solution,water soluble polyethers such as polyethyene glycol, polyvinyls, such aspolyvinyl alcohol and povidone, cellulose derivatives such asmethylcellulose and hydroxypropyl methylcellulose, petroleum derivativessuch as mineral oil and white petrolatum, animal fats such as lanolin,polymers of acrylic acid such as carboxypolymethylene gel, vegetablefats such as peanut oil and polysaccharides such as dextrans, andglycosaminoglycans such as sodium hyaluronate. In some embodiments,additives ordinarily used in the eye drops can be added. Such additivesinclude isotonizing agents (e.g., sodium chloride, etc.), buffer agent(e.g., boric acid, sodium monohydrogen phosphate, sodium dihydrogenphosphate, etc.), preservatives (e.g., benzalkonium chloride,benzethonium chloride, chlorobutanol, etc.), thickeners (e.g.,saccharide such as lactose, mannitol, maltose, etc.; e.g., hyaluronicacid or its salt such as sodium hyaluronate, potassium hyaluronate,etc.; e.g., mucopolysaccharide such as chondroitin sulfate, etc.; e.g.,sodium polyacrylate, carboxyvinyl polymer, crosslinked polyacrylate,polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose,hydroxy propyl cellulose or other agents known to those skilled in theart).

In some cases, the colloid particles include at least one cationic agentand at least one non-ionic surfactant such as a poloxamer, tyloxapol, apolysorbate, a polyoxyethylene castor oil derivative, a sorbitan ester,or a polyoxyl stearate. In some cases, the cationic agent is analkylamine, a tertiary alkyl amine, a quaternary ammonium compound, acationic lipid, an amino alcohol, a biguanidine salt, a cationiccompound or a mixture thereof. In some cases, the cationic agent is abiguanidine salt such as chlorhexidine, polyaminopropyl biguanidine,phenformin, alkylbiguanidine, or a mixture thereof. In some cases, thequaternary ammonium compound is a benzalkonium halide, lauralkoniumhalide, cetrimide, hexadecyltrimethylammonium halide,tetradecyltrimethylammonium halide, dodecyltrimethylammonium halide,cetrimonium halide, benzethonium halide, benzalkonium halide,cetalkonium halide, cetethyldimonium halide, cetylpyridinium halide,benzododecinium halide, chlorallyl methenamine halide, myristalkoniumhalide, stearalkonium halide or a mixture of two or more thereof. Insome cases, cationic agent is a benzalkonium chloride, lauralkoniumchloride, benzododecinium bromide, benzethenium chloride,hexadecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide,dodecyltrimethylammonium bromide or a mixture of two or more thereof. Insome cases, the oil phase is mineral oil and light mineral oil, mediumchain triglycerides (MCT), coconut oil; hydrogenated oils comprisinghydrogenated cottonseed oil, hydrogenated palm oil, hydrogenate castoroil or hydrogenated soybean oil; polyoxyethylene hydrogenated castor oilderivatives comprising poluoxyl-40 hydrogenated castor oil, polyoxyl-60hydrogenated castor oil or polyoxyl-100 hydrogenated castor oil.

5.4.1.6 Formulations for Controlled Release Administration

In some embodiments, provided herein are pharmaceutical compositions forcontrolled release administration containing a compound as providedherein, and a pharmaceutical excipient suitable for controlled releaseadministration. In some embodiments, provided herein are pharmaceuticalcompositions for controlled release administration containing: (i) aneffective amount of a disclosed compound; optionally (ii) an effectiveamount of one or more second agents; and (iii) one or morepharmaceutical excipients suitable for controlled releaseadministration. In some embodiments, the pharmaceutical compositionfurther contains: (iv) an effective amount of a third agent.

Active agents such as the compounds provided herein can be administeredby controlled release means or by delivery devices that are well knownto those of ordinary skill in the art. Examples include, but are notlimited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899;3,536,809; 3,598,123; and U.S. Pat. Nos. 4,008,719; 5,674,533;5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556;5,639,480; 5,733,566; 5,739,108; 5,891,474; 5,922,356; 5,972,891;5,980,945; 5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350;6,248,363; 6,264,970; 6,267,981; 6,376,461; 6,419,961; 6,589,548;6,613,358; 6,699,500 each of which is incorporated herein by reference.Such dosage forms can be used to provide slow or controlled release ofone or more active agents using, for example, hydropropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmoticsystems, multilayer coatings, microparticles, liposomes, microspheres,or a combination thereof to provide the desired release profile invarying proportions. Suitable controlled release formulations known tothose of ordinary skill in the art, including those described herein,can be readily selected for use with the active agents provided herein.Thus, the pharmaceutical compositions provided encompass single unitdosage forms suitable for oral administration such as, but not limitedto, tablets, capsules, gelcaps, and caplets that are adapted forcontrolled release.

All controlled release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non controlledcounterparts. In some embodiments, the use of a controlled releasepreparation in medical treatment is characterized by a minimum of drugsubstance being employed to cure or control the disease, disorder, orcondition in a minimum amount of time. Advantages of controlled releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased subject compliance. In addition, controlledrelease formulations can be used to affect the time of onset of actionor other characteristics, such as blood levels of the drug, and can thusaffect the occurrence of side (e.g., adverse) effects.

In some embodiments, controlled release formulations are designed toinitially release an amount of a compound as provided herein thatpromptly produces the desired therapeutic effect, and gradually andcontinually release other amounts of the compound to maintain this levelof therapeutic or prophylactic effect over an extended period of time.In order to maintain this constant level of the compound in the body,the compound should be released from the dosage form at a rate that willreplace the amount of drug being metabolized and excreted from the body.Controlled release of an active agent can be stimulated by variousconditions including, but not limited to, pH, temperature, enzymes,water, or other physiological conditions or compounds.

In certain embodiments, the pharmaceutical composition can beadministered using intravenous infusion, an implantable osmotic pump, atransdermal patch, liposomes, or other modes of administration. In oneembodiment, a pump can be used (see, Sefton, CRC Crit. Ref. Biomed. Eng.14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., NEngl. J. Med. 321:574 (1989)). In another embodiment, polymericmaterials can be used. In yet another embodiment, a controlled releasesystem can be placed in a subject at an appropriate site determined by apractitioner of skill, i.e., thus requiring only a fraction of thesystemic dose (see, e.g., Goodson, Medical Applications of ControlledRelease, 115-138 (vol. 2, 1984). Other controlled release systems arediscussed in the review by Langer, Science 249:1527-1533 (1990). The oneor more active agents can be dispersed in a solid inner matrix, e.g.,polymethylmethacrylate, polybutylmethacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethyleneterephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinylalcohol andcross-linked partially hydrolyzed polyvinyl acetate, that is surroundedby an outer polymeric membrane, e.g., polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinylchloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer, that is insoluble in body fluids.The one or more active agents then diffuse through the outer polymericmembrane in a release rate controlling step. The percentage of activeagent in such parenteral compositions is highly dependent on thespecific nature thereof, as well as the needs of the subject.

5.4.2 Dosages

A compound described herein can be delivered in the form ofpharmaceutically acceptable compositions which comprise atherapeutically effective amount of one or more compounds describedherein and/or one or more additional therapeutic agents such as achemotherapeutic, formulated together with one or more pharmaceuticallyacceptable excipients. In some instances, the compound described hereinand the additional therapeutic agent are administered in separatepharmaceutical compositions and can (e.g., because of different physicaland/or chemical characteristics) be administered by different routes(e.g., one therapeutic is administered orally, while the other isadministered intravenously). In other instances, the compound describedherein and the additional therapeutic agent can be administeredseparately, but via the same route (e.g., both orally or bothintravenously). In still other instances, the compound described hereinand the additional therapeutic agent can be administered in the samepharmaceutical composition.

The selected dosage level will depend upon a variety of factorsincluding, for example, the activity of the particular compoundemployed, the route of administration, the time of administration, therate of excretion or metabolism of the particular compound beingemployed, the rate and extent of absorption, the duration of thetreatment, other drugs, compounds and/or materials used in combinationwith the particular compound employed, the age, sex, weight, condition,general health and prior medical history of the patient being treated,and like factors well known in the medical arts.

In general, a suitable daily dose of a compound described herein and/ora chemotherapeutic will be that amount of the compound which, in someembodiments, can be the lowest dose effective to produce a therapeuticeffect. Such an effective dose will generally depend upon the factorsdescribed above. Generally, doses of the compounds described herein fora patient, when used for the indicated effects, will range from about0.0001 mg to about 100 mg per day, or about 0.001 mg to about 100 mg perday, or about 0.01 mg to about 100 mg per day, or about 0.1 mg to about100 mg per day, or about 0.0001 mg to about 500 mg per day, or about0.001 mg to about 500 mg per day, or about 0.01 mg to 1000 mg, or about0.01 mg to about 500 mg per day, or about 0.1 mg to about 500 mg perday, or about 1 mg to 50 mg per day, or about 5 mg to 40 mg. Anexemplary dosage is about 10 to 30 mg per day. In some embodiments, fora 70 kg human, a suitable dose would be about 0.05 to about 7 g/day,such as about 0.05 to about 2.5 g/day. Actual dosage levels of theactive ingredients in the pharmaceutical compositions described hereincan be varied so as to obtain an amount of the active ingredient whichis effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient. In some instances, dosage levels below thelower limit of the aforesaid range can be more than adequate, while inother cases still larger doses can be employed without causing anyharmful side effect, e.g., by dividing such larger doses into severalsmall doses for administration throughout the day.

In some embodiment, the daily dose of a compound described herein canrange from about 0.0001 mg/kg to about 1000 mg/kg, about 0.001 mg/kg toabout 1000 mg/kg, about 0.01 mg/kg to about 1000 mg/kg, about 0.1 mg/kgto about 1000 mg/kg, about 0.0001 mg/kg to about 500 mg/kg, about 0.001mg/kg to about 500 mg/kg, about 0.01 mg/kg to 100 mg/kg, about 0.01mg/kg to about 100 mg/kg, about 0.1 mg/kg to about 100 mg/kg, about 0.01mg/kg to 50 mg/kg, about 0.05 mg/kg to 20 mg/kg, or about 0.05 mg/kg to10 mg/kg. For example, the daily dose can be about 10 mg/kg, 5 mg/kg,1.5 mg/kg, 0.5 mg/kg, 0.15 mg/kg, or about 0.05 mg/kg For example, thedaily dose can be about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about2.5 mg/kg, about 3 mg/kg, about 3.1 mg/kg, about 3.2 mg/kg, about 3.3mg/kg, about 3.4 mg/kg, about 3.5 mg/kg, about 4 mg/kg, about 5 mg/kg,about 6 mg/kg, about 7 mg/kg, about 8 mg/kg, about 9 mg/kg, or about 10mg/kg.

In some embodiments, the compounds can be administered daily, everyother day, three times a week, twice a week, weekly, or bi-weekly. Thedosing schedule can include a “drug holiday,” e.g., the drug can beadministered for two weeks on, one week off, or three weeks on, one weekoff, or four weeks on, one week off, etc., or continuously, without adrug holiday. The compounds can be administered orally, intravenously,intraperitoneally, topically, transdermally, intramuscularly,subcutaneously, intranasally, sublingually, or by any other route.

In some embodiments, a compound as provided herein is administered inmultiple doses. Dosing can be about once, twice, three times, fourtimes, five times, six times, or more than six times per day. Dosing canbe about once a month, about once every two weeks, about once a week, orabout once every other day. In another embodiment, a compound asprovided herein and another agent are administered together from aboutonce per day to about 6 times per day. In another embodiment, theadministration of a compound as provided herein and an agent continuesfor less than about 7 days. In yet another embodiment, theadministration continues for more than about 6 days, about 10 days,about 14 days, about 28 days, about two months, about six months, orabout one year. In some cases, continuous dosing is achieved andmaintained as long as necessary.

Administration of the pharmaceutical compositions as provided herein cancontinue as long as necessary. In some embodiments, an agent as providedherein is administered for more than about 1, about 2, about 3, about 4,about 5, about 6, about 7, about 14, about 21, or about 28 days. In someembodiments, an agent as provided herein is administered for less thanabout 28, about 21, about 14, about 7, about 6, about 5, about 4, about3, about 2, or about 1 day. In some embodiments, an agent as providedherein is administered for about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 14, about 21, or about 28 days. In someembodiments, an agent as provided herein is administered chronically onan ongoing basis, e.g., for the treatment of chronic effects.

Since the compounds described herein can be administered in combinationwith other treatments (such as additional chemotherapeutics, radiationor surgery), the doses of each agent or therapy can be lower than thecorresponding dose for single-agent therapy. The dose for single-agenttherapy can range from, for example, about 0.0001 to about 200 mg, orabout 0.001 to about 100 mg, or about 0.01 to about 100 mg, or about 0.1to about 100 mg, or about 1 to about 50 mg per kilogram of body weightper day.

When a compound provided herein, is administered in a pharmaceuticalcomposition that comprises one or more agents, and the agent has ashorter half-life than the compound provided herein unit dose forms ofthe agent and the compound provided herein can be adjusted accordingly.

5.4.3 Kits

In some embodiments, provided herein are kits. The kits can include acompound or pharmaceutical composition as described herein, in suitablepackaging, and written material that can include instructions for use,discussion of clinical studies, listing of side effects, and the like.Such kits can also include information, such as scientific literaturereferences, package insert materials, clinical trial results, and/orsummaries of these and the like, which indicate or establish theactivities and/or advantages of the pharmaceutical composition, and/orwhich describe dosing, administration, side effects, drug interactions,or other information useful to the health care provider. Suchinformation can be based on the results of various studies, for example,studies using experimental animals involving in vivo models and studiesbased on human clinical trials.

In some embodiments, a memory aid is provided with the kit, e.g., in theform of numbers next to the tablets or capsules whereby the numberscorrespond with the days of the regimen which the tablets or capsules sospecified should be ingested. Another example of such a memory aid is acalendar printed on the card, e.g., as follows “First Week, Monday,Tuesday, . . . etc. . . . Second Week, Monday, Tuesday, . . . etc.”Other variations of memory aids will be readily apparent. A “daily dose”can be a single tablet or capsule or several tablets or capsules to betaken on a given day.

The kit can further contain another agent. In some embodiments, thecompound as provided herein and the agent are provided as separatepharmaceutical compositions in separate containers within the kit. Insome embodiments, the compound as provided herein and the agent areprovided as a single pharmaceutical composition within a container inthe kit. Suitable packaging and additional articles for use (e.g.,measuring cup for liquid preparations, foil wrapping to minimizeexposure to air, and the like) are known in the art and can be includedin the kit. In other embodiments, kits can further comprise devices thatare used to administer the active agents. Examples of such devicesinclude, but are not limited to, syringes, drip bags, patches, andinhalers. Kits described herein can be provided, marketed and/orpromoted to health providers, including physicians, nurses, pharmacists,formulary officials, and the like. Kits can also, in some embodiments,be marketed directly to the consumer.

An example of such a kit is a so-called blister pack. Blister packs arewell known in the packaging industry and are being widely used for thepackaging of pharmaceutical unit dosage forms (tablets, capsules, andthe like). Blister packs generally consist of a sheet of relativelystiff material covered with a foil of a preferably transparent plasticmaterial. During the packaging process, recesses are formed in theplastic foil. The recesses have the size and shape of the tablets orcapsules to be packed. Next, the tablets or capsules are placed in therecesses and the sheet of relatively stiff material is sealed againstthe plastic foil at the face of the foil which is opposite from thedirection in which the recesses were formed. As a result, the tablets orcapsules are sealed in the recesses between the plastic foil and thesheet. The strength of the sheet is such that the tablets or capsulescan be removed from the blister pack by manually applying pressure onthe recesses whereby an opening is formed in the sheet at the place ofthe recess. The tablet or capsule can then be removed via said opening.

Kits can further comprise pharmaceutically acceptable vehicles that canbe used to administer one or more active agents. For example, if anactive agent is provided in a solid form that must be reconstituted forparenteral administration, the kit can comprise a sealed container of asuitable vehicle in which the active agent can be dissolved to form aparticulate-free sterile solution that is suitable for parenteraladministration. Examples of pharmaceutically acceptable vehiclesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

The present disclosure further encompasses anhydrous pharmaceuticalcompositions and dosage forms comprising an active ingredient, sincewater can facilitate the degradation of some compounds. For example,water can be added (e.g., about 5%) in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. Anhydrous pharmaceutical compositions and dosage forms can beprepared using anhydrous or low moisture containing ingredients and lowmoisture or low humidity conditions. For example, pharmaceuticalcompositions and dosage forms which contain lactose can be madeanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected. An anhydrouspharmaceutical composition can be prepared and stored such that itsanhydrous nature is maintained. Accordingly, anhydrous pharmaceuticalcompositions can be packaged using materials known to prevent exposureto water such that they can be included in suitable formulary kits.Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastic or the like, unit dose containers,blister packs, and strip packs.

5.5. Therapeutic Methods

Provided herein is a method for treating a PI3K mediated disorder in asubject, comprising administering a therapeutically effective amount ofa solid form provided herein, or a pharmaceutical composition providedherein to said subject. In one embodiment, the PI3K mediated disorder iscancer, an inflammatory disease or an auto-immune disease. In oneembodiment, the cancer is solid tumor.

Phosphoinositide 3-kinases (PI3Ks) are members of a conserved family oflipid kinases that regulate numerous cell functions, includingproliferation, differentiation, cell survival and metabolism. Severalclasses of PI3Ks exist in mammalian cells, including Class IA subgroup(e.g., PI3K-α, β, δ), which are generally activated by receptor tyrosinekinases (RTKs); Class IB (e.g., PI3K-γ), which is activated by G-proteincoupled receptors (GPCRs), among others. PI3Ks exert their biologicalactivities via a “PI3K-mediated signaling pathway” that includes severalcomponents that directly and/or indirectly transduce a signal triggeredby a PI3K, including the generation of second messengerphophotidylinositol, 3,4,5-triphosphate (PIP3) at the plasma membrane,activation of heterotrimeric G protein signaling, and generation offurther second messengers such as cAMP, DAG, and IP3, all of which leadsto an extensive cascade of protein kinase activation (reviewed inVanhaesebroeck, B. et al. (2001) Annu Rev Biochem. 70:535-602). Forexample, PI3K-δ is activated by cellular receptors through interactionbetween the PI3K regulatory subunit (p85) SH2 domains, or through directinteraction with RAS. PIP3 produced by PI3K activates effector pathwaysdownstream through interaction with plextrin homology (PH) domaincontaining enzymes (e.g., PDK-1 and AKT [PKB]). (Fung-Leung W P. (2011)Cell Signal. 23(4):603-8). Unlike PI3K-δ, PI3K-γ is not associated witha regulatory subunit of the p85 family, but rather with a regulatorysubunit in the p101 or p84 families. PI3K-γ is associated with GPCRs,and is responsible for the very rapid induction of PIP3. PI3K-γ can bealso activated by RAS.

In some embodiments, provided herein are methods of modulating a PI3kinase activity (e.g., selectively modulating) by contacting the kinasewith an effective amount of a compound as provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein. Modulation can be inhibition (e.g., reduction) or activation(e.g., enhancement) of kinase activity. In some embodiments, providedherein are methods of inhibiting kinase activity by contacting thekinase with an effective amount of a compound as provided herein insolution. In some embodiments, provided herein are methods of inhibitingthe kinase activity by contacting a cell, tissue, organ that express thekinase of interest, with a compound provided herein. In someembodiments, provided herein are methods of inhibiting kinase activityin a subject by administering into the subject an effective amount of acompound as provided herein, or a pharmaceutically acceptable formthereof. In some embodiments, the kinase activity is inhibited (e.g.,reduced) by more than about 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%,when contacted with a compound provided herein as compared to the kinaseactivity without such contact. In some embodiments, provided herein aremethods of inhibiting PI3 kinase activity in a subject (includingmammals such as humans) by contacting said subject with an amount of acompound as provided herein sufficient to inhibit or reduce the activityof the PI3 kinase in said subject.

In some embodiments, the kinase is a lipid kinase or a protein kinase.In some embodiments, the kinase is selected from a PI3 kinase includingdifferent isoforms, such as PI3 kinase α, PI3 kinase β, PI3 kinase γ,PI3 kinase δ; DNA-PK; mTOR; Abl, VEGFR, Ephrin receptor B4 (EphB4); TEKreceptor tyrosine kinase (TIE2); FMS-related tyrosine kinase 3 (FLT-3);Platelet derived growth factor receptor (PDGFR); RET; ATM; ATR; hSmg-1;Hck; Src; Epidermal growth factor receptor (EGFR); KIT; Insulin Receptor(IR); and IGFR.

As used herein, a “PI3K-mediated disorder” refers to a disease orcondition involving aberrant PI3K-mediated signaling pathway. In oneembodiment, provided herein is a method of treating a PI3K mediateddisorder in a subject, the method comprising administering atherapeutically effective amount of a compound as provided herein, or apharmaceutically acceptable form thereof, or a pharmaceuticalcomposition as provided herein. In some embodiments, provided herein isa method of treating a PI3K-δ or PI3K-γ mediated disorder in a subject,the method comprising administering a therapeutically effective amountof a compound as provided herein, or a pharmaceutically acceptable formthereof, or a pharmaceutical composition as provided herein. In someembodiments, provided herein is a method for inhibiting at least one ofPI3K-δ and PI3K-γ, the method comprising contacting a cell expressingPI3K in vitro or in vivo with an effective amount of a compound orcomposition provided herein. PI3Ks have been associated with a widerange of conditions, including immunity, cancer and thrombosis (reviewedin Vanhaesebroeck, B. et al. (2010) Current Topics in Microbiology andImmunology, DOI 10.1007/82_2010_65). For example, Class I PI3Ks,particularly PI3K-γ and PI3K-δ isoforms, are highly expressed inleukocytes and have been associated with adaptive and innate immunity;thus, these PI3Ks are believed to be important mediators in inflammatorydisorders and hematologic malignancies (reviewed in Harris, S J et al.(2009) Curr Opin Investig Drugs 10(11):1151-62); Rommel C. et al. (2007)Nat Rev Immunol 7(3):191-201; Durand C A et al. (2009) J Immunol.183(9):5673-84; Dil N, Marshall A J. (2009) Mol Immunol. 46(10):1970-8;Al-Alwan M M et al. (2007) J Immunol. 178(4):2328-35; Zhang T T, et al.(2008) J Allergy Clin Immunol. 2008; 122(4):811-819.e2; Srinivasan L, etal. (2009) Cell 139(3):573-86).

PI3K-γ Activities

PI3K-γ is a Class 1B PI3K that associates with the p101 and p84(p87PIKAP) adaptor proteins, and canonically signals through GPCRs.Non-canonical activation through tyrosine kinase receptors and RAS canoccur. Activated PI3K-γ leads to production of PIP3, which serves as adocking site for downstream effector proteins including AKT and BTK,bringing these enzymes to the cell membrane where they may be activated.A scaffolding role for PI3K-γ has been proposed and may contribute tothe activation of the RAS/MEK/ERK pathway. The interaction with the RASpathway explains activities attributed to kinase dead PI3K-γ in cells orin animals. PI3K-γ is essential for function of a variety of immunecells and pathways. Chemokine responses (including IL-8, fMLP, and C5a),leading to neutrophil, basophil or monocyte cell migration, is dependenton PI3K-γ (HIRSCH et al., “Central Role for G Protein-CoupledPhosphoinositide 3-Kinase γ in Inflammation,” Science 287:1049-1053(2000); SASAKI et al., “Function of PI3Kγ in Thymocyte Development, TCell Activation, and Neutrophil Migration,” Science 287:1040-1046(2000); LI et al., “Roles of PLC-β2 and -β3 and PI3Kγ inChemoattractant-Mediated Signal Transduction,” Science 287:1046-1049(2000)). The requirement for PI3K-γ-dependent neutrophil migration isdemonstrated by failure of arthritis development in the K/BXN serumtransfer arthritis model in PI3K-γ knockout mice (Randis et al., Eur. J.Immunol., 2008, 38(5), 1215-24). Similarly, the mice fail to developcellular inflammation and airway hyper-responsiveness in the ovalbumininduced asthma model (Takeda et al., J. Allergy Clin. Immunol., 2009;123, 805-12). PI3K-γ deficient mice also have defects in T-helper cellfunction. T-cell cytokine production and proliferation in response toactivation is reduced, and T helper dependent viral clearance isdefective (Sasaki et al., Science, 2000, 287, 1040-46). T cell dependentinflammatory disease models including EAE also do not develop in PI3K-γdeficient mice, and both the T-cell activation defect and cellularmigration defects may contribute to efficacy in this model (Comerfold,PLOS One, 2012, 7, e45095). The imiquimod psoriasis model has also beenused to demonstrate the importance of PI3K-γ in the inflammatoryresponse. Using PI3K-γ deficient mice in this model, the accumulation ofγδ T cells in the skin is blocked, as well as dendritic cell maturationand migration (ROLLER et al., “Blockade of Phosphatidylinositol 3-Kinase(PI3K)δ or PI3Kγ Reduces IL-17 and Ameliorates Imiquimod-InducedPsoriasis-like Dermatitis,” J. Immunol. 189:4612-4620 (2012)). The roleof PI3K-γ in cellular trafficking can also be demonstrated in oncologymodels where tumor inflammation is important for growth and metastasisof cancers. In the Lewis Lung Carcinoma model, monocyte activation,migration, and differentiation in tumors are defective. This defectresults in a reduction in tumor growth and extended survival in PI3K-γdeficient mice (Schmid et al., Cancer Cell, 2011, 19, 715-27) or upontreatment with inhibitors that target PI3K-γ. In pancreatic cancer,PI3K-γ can be inappropriately expressed, and in this solid tumor canceror others where PI3K-γ plays a functional role, inhibition of PI3K-γ canbe beneficial.

For instance, while not wishing to be bound by theory, PI3K-γ isexpressed in Gr1+CD11b+myeloid cells, and directly promotes myeloid cellinvasion and consequently, immunosuppression of pancreatic ductalcarcinomas. Hardamon et. al., Proceedings: AACR 103rd Annual Meeting2012, Cancer Research: Apr. 15, 2012; Volume 72, Issue 8, Supplement 1Inhibition of PI3K-γ also shows promise for the treatment of hematologicmalignancies. In a T-ALL model employing a T cell directed knockout ofpten, PI3K-δ and PI3K-γ are both essential for the appropriatedevelopment of disease, as shown with genetic deletion of both genes(Subramaniam et al. Cancer Cell 21, 459-472, 2012). In addition, in thisT-ALL model, treatment with a small molecule inhibitor of both kinasesleads to extended survival of these mice. In CLL, chemokine networkssupport a pseudo-follicular microenvironment that includes Nurse likecells, stromal cells and T-helper cells. The roles of PI3K-γ in thenormal chemokine signaling and T cell biology suggest the value ofinhibiting this target in CLL (BURGER, “Inhibiting B-Cell ReceptorSignaling Pathways in Chronic Lymphocytic Leukemia,” Curr. Mematol.Malig. Rep. 7:26-33 (2012)). Accordingly, PI3K-γ inhibitors aretherapeutically interesting for diseases of the immune system where celltrafficking and T cell or myeloid cell function is important. Inoncology, solid tumors that are dependent on tumor inflammation, ortumors with high levels of PI3K-γ expression, can be targeted. Forhematological cancers, a special role for PI3K-γ and PI3K-δ isoforms inTALL and potentially in CLL suggests targeting these PI3Ks in thesediseases.

Without being limited by a particular theory, PI3K-γ has been shown toplay roles in inflammation, arthritis, asthma, allergy, multiplesclerosis (MS), and cancer, among others (e.g., Ruckle et al., NatureRev., Drug Discovery, 2006, 5, 903-18; Schmid et al., “Myeloid cells intumor inflammation,” Vascular Cell, 2012, doi:10.118612045-824X-4-14).For example, PI3K-γ functions in multiple signaling pathways involved inleukocyte activation and migration. PI3K-γ has been shown to drivepriming and survival of autoreactive CD4⁺ T cells during experimentalautoimmune encephalomyelitis (EAE), a model for MS. When administeredfrom onset of EAE, a PI3K-γ inhibitor has been shown to cause inhibitionand reversal of clinical disease, and reduction of demyelination andcellular pathology in the CNS (Comerford et al., PLOS One, 2012, 7,e45095). PI3K-γ also regulates thymocyte development, T cell activation,neutrophil migration, and the oxidative burst (Sasaki et al., Science,2000, 287, 1040-46). In addition, it is shown that allergic airwayhyper-responsiveness, inflammation, and remodeling do not develop inPI3K-γ deficient mice (Takeda et al., J. Allergy Clin. Immunol., 2009;123, 805-12). PI3K-γ is shown to be required for chemoattractant-inducedproduction of phosphatidylinositol 3,4,5-trisphosphate and has animportant role in chemoattractant-induced superoxide production andchemotaxis in mouse neutrophils and in production of T cell-independentantigen-specific antibodies composed of the immunoglobulin λ light chain(Li et al., Science, 2000, 287, 1046-49). PI3K-γ is reported to be acrucial signaling molecule required for macrophage accumulation ininflammation (Hirsch et al., Science, 2000, 287, 1049-53). In cancers,pharmacological or genetic blockade of p110γ suppresses inflammation,growth, and metastasis of implanted and spontaneous tumors, suggestingthat PI3K-γ can be an important therapeutic target in oncology (Schmidet al., Cancer Cell, 2011, 19, 715-27). For example, it is shown thatPI3K-γ has a tumor-specific high accumulation in pancreatic ductaladenocarcinoma (PDAC) in human, signifying a role of PI3K-γ inpancreatic cancer (Edling et al., Human Cancer Biology, 2010, 16(2),4928-37).

In certain embodiments, provided herein are methods of treating orpreventing a PI3K-gamma mediated disorder in a subject, comprisingadministering to the subject a therapeutically effective amount of acompound provided herein, e.g., a selective PI3K-γ inhibitor, e.g.,Compound 1), or a pharmaceutically acceptable form thereof.

In one embodiment, the subject has or is at risk of having a PI3K-gammamediated disorder selected from cancer, an inflammatory disease, or anautoimmune disease. In one embodiment, the cancer is a solid tumor. Inone embodiment, the cancer is selected from one or more of: a cancer ofthe pulmonary system, a brain cancer, a cancer of the gastrointestinaltract, a skin cancer, a genitourinary cancer, a pancreatic cancer, alung cancer, a medulloblastoma, a basal cell carcinoma, a glioma, abreast cancer, a prostate cancer, a testicular cancer, an esophagealcancer, a hepatocellular cancer, a gastric cancer, a gastrointestinalstromal tumor (GIST), a colon cancer, a colorectal cancer, an ovariancancer, a melanoma, a neuroectodermal tumor, head and neck cancer, asarcoma, a soft-tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma,a chondrosarcoma, an osteogenic sarcoma, a chordoma, an angiosarcoma, anendotheliosarcoma, a lymphangiosarcoma, a lymphangioendotheliosarcoma, asynovioma, a mesothelioma, a leiomyosarcoma, a cervical cancer, auterine cancer, an endometrial cancer, a carcinoma, a bladder carcinoma,an epithelial carcinoma, a squamous cell carcinoma, an adenocarcinoma, abronchogenic carcinoma, a renal cell carcinoma, a hepatoma, a bile ductcarcinoma, a neuroendocrine cancer, a carcinoid tumor, diffuse typegiant cell tumor, and glioblastoma.

In one embodiment, the cancer is a hematological cancer.

In one embodiment, the inflammatory disease is arthritis.

In one embodiment, the subject is a human. In one embodiment, thesubject is identified as having or being at risk of having a PI3K-gammamediated disorder via the use of a biomarker.

In one embodiment, the therapeutically effective dose is about 2 mg,about 1-3 mg, about 1-5 mg, about 1-10 mg, about 0.5-20 mg, about 0.1-50mg per day, about 0.1-75 mg per day, about 0.1-100 mg per day, about0.1-250 mg per day, about 0.1-500 mg per day, about 0.1-1000 mg per day,about 1-50 mg per day, about 1-75 mg per day, about 1-100 mg per day,about 1-250 mg per day, about 1-500 mg per day, about 1-1000 mg per day,about 10-50 mg per day, about 10-75 mg per day, about 10-100 mg per day,about 10-250 mg per day, about 10-500 mg per day, about 10-1000 mg perday, about 100-500 mg per day, or about 100-1000 mg per day. In oneembodiment, the therapeutically effective dose is about 0.029 mg/kg,about 0.014-0.14 mg/kg, about 0.02-0.04 mg/kg, about 0.01-0.05 mg/kg,about 0.01-0.1, or about 0.01-0.5 mg/kg. In one embodiment, the compoundis administered once every two days. In one embodiment, wherein thecompound is administered once per day. In one embodiment, the compoundis administered twice per day.

In one embodiment, the compound is administered at a dose such that thelevel of the compound in the subject is higher than the compound's IC50of PI3K-gamma inhibition during at least 70%, 80%, 90%, 95%, 97%, 98%,or 99% of a selected time period, e.g., 6 hours, 12 hours, 24 hours, or48 hours immediately following the administration. In one embodiment,the compound is administered at a dose such that the level of thecompound in the subject is higher than the compound's IC90 of PI3K-gammainhibition during at least 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, or99% of a selected time period, e.g., 6 hours, 12 hours, 24 hours, or 48hours, immediately following the administration. In one embodiment, thecompound is administered at a dose such that the level of the compoundin the subject does not rise higher than the compound's IC20 or IC50 ofPI3K-delta inhibition within a selected time period, e.g., 6 hours, 12hours, 24 hours, or 48 hours, immediately following the administration.In one embodiment, the level of the compound is measured from thesubject's plasma. In one embodiment, the level of the compound ismeasured from the subject's tissue. In one embodiment, the compound isadministered at a dose such that it provides at least 50% inhibition ofPI3K-gamma in the subject but less than 10% or 20% inhibition ofPI3K-delta in the subject.

In one embodiment, the subject is a human and the compound has ahalf-life of about 10-13 hours in the subject. In one embodiment, themethod further comprises administering to the subject a secondtherapeutic agent that is a P-gp substrate. In one embodiment, thesecond therapeutic agent is Norvir (ritonavir).

PI3K-δ and/or PI3K-γ Activities

PI3K-δ has roles in impairments of B-cell signaling and development,antibody production, T-cell function, Th1 and Th2 differentiation, andmast and basophil degranulation. Without being limited by a particulartheory, PI3K-γ has roles in T-cell function, neutrophil and macrophagerecruitment, macrophage activation, neutrophil oxidative burst, anddendritic cell migration Inhibition of PI3K-δ and/or PI3K-γ isoforms canresult in efficacy against inflammation and cancer, e.g., in arthritis,asthma, multiple sclerosis (MS), and tumor models. For example,deficiency in PI3K-δ and/or PI3K-γ can result in efficacy in K/B×Narthritis model (Kyburz et al., Springer Semin. Immunopathology, 2003,25, 79-90) or K/B×N serum transfer model of arthritis (Randis et al.,Eur. J. Immunol., 2008, 38(5), 1215-24), where it is shown thatrecognition of the immune complexes depends on both PI3K-δ and PI3K-γ,whereas cell migration is dependent on PI3K-γ. Deficiency in PI3K-δ orPI3K-γ can also result in efficacy in murine ovalbumin (OVA) inducedallergic asthma model (Lee et al., FASEB J., 2006, 20, 455-65; Takeda etal., J. Allergy Clin. Immunol., 2009; 123, 805-12), where it is shownthat inhibition of either PI3K-δ or PI3K-γ inhibits ovalbumin inducedlung infiltration and improves airway responsiveness. Deficiency inPI3K-δ or PI3K-γ can also result in efficacy in murine experimentalautoimmune encephalomyelitis (model for MS), where it is shown thatPI3K-γ deletion may provide better efficacy as compared to PI3K-δdeletion (Haylock-Jacob et al., J. Autoimmunity, 2011, 36, 278-87;Comerford et al., PLOS One, 2012, 7, e45095), including reduction inT-cell receptor induced CD4⁺ T cell activation, leukocyte infiltrationand Th1/Th17 responses, and dendritic cell migration (Comerfold, PLOSOne, 2012, 7, e45095). Furthermore, inhibition of PI3K-γ can also resultin decreased tumor inflammation and growth (e.g., Lewis lung carcinomamodel, Schmid et al., Cancer Cell, 2011, 19(6), 715-27). PI3K-γ deletioncombined with PI3K-δ deletion results in increased survival in T-cellacute lymphoblastic leukemia (T-ALL) (Subramaniam et al., Cancer Cell,2012, 21, 459-72). Inhibitors of both PI3K-δ and PI3K-γ are also shownto be efficacious in PTEN-deleted T-ALL cell line (MOLT-4). In theabsence of PTEN phosphatase tumor suppressor function, PI3K-δ or PI3K-γalone can support the development of leukemia, whereas inactivation ofboth isoforms suppresses tumor formation. Thus, inhibitors of PI3K-δand/or PI3K-γ can be useful in treating inflammation, such as arthritis,allergic asthma, and MS; and in treating cancer, for example, due toeffects such as reductions in solid tumor associated inflammation,angiogenesis and tumor progression.

The importance of PI3K-δ in the development and function of B-cells issupported from inhibitor studies and genetic models. PI3K-δ is animportant mediator of B-cell receptor (BCR) signaling, and is upstreamof AKT, calcium flux, PLCγ, MAP kinase, P70S6k, and FOXO3a activation.PI3K-δ is also important in IL4R, S1P, and CXCR5 signaling, and has beenshown to modulate responses to toll-like receptors 4 and 9 Inhibitors ofPI3K-δ have shown the importance of PI3K-δ in B-cell development(Marginal zone and B1 cells), B-cell activation, chemotaxis, migrationand homing to lymphoid tissue, and in the control of immunoglobulinclass switching leading to the production of IgE. Clayton E et al.(2002) J Exp Med. 196(6):753-63; Bilancio A, et al. (2006) Blood107(2):642-50; Okkenhaug K. et al. (2002) Science 297(5583):1031-4;Al-Alwan M M et al. (2007) J Immunol. 178(4):2328-35; Zhang T T, et al.(2008) J Allergy Clin Immunol. 2008; 122(4):811-819.e2; Srinivasan L, etal. (2009) Cell 139(3):573-86).

In T-cells, PI3K-δ has been demonstrated to have a role in T-cellreceptor and cytokine signaling, and is upstream of AKT, PLCγ, andGSK3b. In PI3K-δ deletion or kinase-dead knock-in mice, or in inhibitorstudies, T-cell defects including proliferation, activation, anddifferentiation have been observed, leading to reduced T helper cell 2(TH2) response, memory T-cell specific defects (DTH reduction), defectsin antigen dependent cellular trafficking, and defects inchemotaxis/migration to chemokines (e.g., S1P, CCR7, CD62L). (Garçon F.et al. (2008) Blood 111(3):1464-71; Okkenhaug K et al. (2006). JImmunol. 177(8):5122-8; Soond D R, et al. (2010) Blood 115(11):2203-13;Reif K, (2004). J Immunol. 2004; 173(4):2236-40; Ji H. et al. (2007)Blood 110(8):2940-7; Webb L M, et al. (2005) J Immunol. 175(5):2783-7;Liu D, et al. (2010) J Immunol. 184(6):3098-105; Haylock-Jacobs S, etal. (2011) J Autoimmun. 2011; 36(3-4):278-87; Jarmin S J, et al. (2008)J Clin Invest. 118(3): 1154-64).

Numerous publications support roles of PI3K-δ and PI3K-γ in thedifferentiation, maintenance, and activation of immune and malignantcells, as described in more detail herein.

PI3K-δ and PI3K-γ isoforms are preferentially expressed in leukocyteswhere they have distinct and non-overlapping roles in immune celldevelopment and function. See, e.g., PURI and GOLD, “Selectiveinhibitors of phosphoinositide 3-kinase delta: modulators of B-cellfunction with potential for treating autoimmune inflammatory disease andB-cell malignancies,” Front. Immunol. 3:256 (2012); BUITENHUIS et al.,“The role of the PI3K-PKB signaling module in regulation ofhematopoiesis,” Cell Cycle 8(4):560-566 (2009); HOELLENRIEGEL andBURGER, “Phosphoinositide 3′-kinase delta: turning off BCR signaling inChronic Lymphocytic Leukemia,” Oncotarget 2(10):737-738 (2011); HIRSCHet al., “Central Role for G Protein-Coupled Phosphoinositide 3-Kinase γin Inflammation,” Science 287:1049-1053 (2000); LI et al., “Roles ofPLC-β2 and -β3 and PI3Kγ in Chemoattractant-Mediated SignalTransduction,” Science 287:1046-1049 (2000); SASAKI et al., “Function ofPI3Kγ in Thymocyte Development, T Cell Activation, and NeutrophilMigration,” Science 287:1040-1046 (2000); CUSHING et al., “PI3Kδ andPI3Kγ as Targets for Autoimmune and Inflammatory Diseases,” J. Med.Chem. 55:8559-8581 (2012); MAXWELL et al., “Attenuation ofphosphoinositide 3-kinase δ signaling restrains autoimmune disease,” J.Autoimmun. 38:381-391 (2012); HAYLOCK-JACOBS et al., “PI3Kδ drives thepathogenesis of experimental autoimmune encephalomyelitis by inhibitingeffector T cell apoptosis and promoting Th17 differentiation,” J.Autoimmun. 36:278-287 (2011); SOOND et al., “PI3K p110δ regulates T-cellcytokine production during primary and secondary immune responses inmice and humans,” Blood 115(11):2203-2213 (2010); ROLLER et al.,“Blockade of Phosphatidylinositol 3-Kinase (PI3K)δ or PI3Kγ ReducesIL-17 and Ameliorates Imiquimod-Induced Psoriasis-like Dermatitis,” J.Immunol. 189:4612-4620 (2012); CAMPS et al., “Blockade of PI3Kγsuppresses joint inflammation and damage in mouse models of rheumatoidarthritis,” Nat. Med. 11(9):936-943 (2005). As key enzymes in leukocytesignaling, PI3K-δ and PI3K-γ facilitate normal B-cell, T-cell andmyeloid cell functions including differentiation, activation, andmigration. See, e.g., HOELLENRIEGEL and BURGER, “Phosphoinositide3′-kinase delta: turning off BCR signaling in Chronic LymphocyticLeukemia,” Oncotarget 2(10):737-738 (2011); CUSHING et al., “PI3Kδ andPI3Kγ as Targets for Autoimmune and Inflammatory Diseases,” J. Med.Chem. 55:8559-8581 (2012). PI3K-δ or PI3K-γ activity is critical forpreclinical models of autoimmune and inflammatory diseases. See, e.g.,HIRSCH et al., “Central Role for G Protein-Coupled Phosphoinositide3-Kinase γ in Inflammation,” Science 287:1049-1053 (2000); LI et al.,“Roles of PLC-β2 and -β3 and PI3Kγ in Chemoattractant-Mediated SignalTransduction,” Science 287:1046-1049 (2000); SASAKI et al., “Function ofPI3Kγ in Thymocyte Development, T Cell Activation, and NeutrophilMigration,” Science 287:1040-1046 (2000); CUSHING et al., “PI3Kδ andPI3Kγ as Targets for Autoimmune and Inflammatory Diseases,” J. Med.Chem. 55:8559-8581 (2012); MAXWELL et al., “Attenuation ofphosphoinositide 3-kinase δ signaling restrains autoimmune disease,” J.Autoimmun. 38:381-391 (2012); HAYLOCK-JACOBS et al., “PI3Kδ drives thepathogenesis of experimental autoimmune encephalomyelitis by inhibitingeffector T cell apoptosis and promoting Th17 differentiation,” J.Autoimmun. 36:278-287 (2011); SOOND et al., “PI3K p110δ regulates T-cellcytokine production during primary and secondary immune responses inmice and humans,” Blood 115(11):2203-2213 (2010); ROLLER et al.,“Blockade of Phosphatidylinositol 3-Kinase (PI3K)δ or PI3Kγ ReducesIL-17 and Ameliorates Imiquimod-Induced Psoriasis-like Dermatitis,” J.Immunol. 189:4612-4620 (2012); CAMPS et al., “Blockade of PI3Kγsuppresses joint inflammation and damage in mouse models of rheumatoidarthritis,” Nat. Med. 11(9):936-943 (2005). Given the key role forPI3K-δ and PI3K-γ in immune function, inhibitors of the PI3K-δ and/or γhave therapeutic potential in immune-related inflammatory or neoplasticdiseases.

PI3K-δ and PI3K-γ are central to the growth and survival of B- andT-cell malignancies and inhibition of these isoforms may effectivelylimit these diseases. See, e.g., SUBRAMANIAM et al., “TargetingNonclassical Oncogenes for Therapy in T-ALL,” Cancer Cell 21:459-472(2012); LANNUTTI et al., “CAL-101 a p110δ selectivephosphatidylinositol-3-kinase inhibitor for the treatment of B-cellmalignancies, inhibits PI3K signaling and cellular viability,” Blood117(2):591-594 (2011). PI3K-δ and PI3K-γ support the growth and survivalof certain B-cell malignancies by mediating intracellular BCR signalingand interactions between the tumor cells and their microenvironment.See, e.g., PURI and GOLD, “Selective inhibitors of phosphoinositide3-kinase delta: modulators of B-cell function with potential fortreating autoimmune inflammatory disease and B-cell malignancies,”Front. Immunol. 3:256 (2012); HOELLENRIEGEL et al., “Thephosphoinositide 3′-kinase delta inhibitor, CAL-101, inhibits B-cellreceptor signaling and chemokine networks in chronic lymphocyticleukemia,” Blood 118(13): 3603-3612 (2011); BURGER, “Inhibiting B-CellReceptor Signaling Pathways in Chronic Lymphocytic Leukemia,” Curr.Mematol. Malig. Rep. 7:26-33 (2012). Increased BCR signaling is acentral pathologic mechanism of B-cell malignancies and PI3K activationis a direct consequence of BCR pathway activation. See, e.g., BURGER,“Inhibiting B-Cell Receptor Signaling Pathways in Chronic LymphocyticLeukemia,” Curr. Mematol. Malig. Rep. 7:26-33 (2012); HERISHANU et al.,“The lymph node microenvironment promotes B-cell receptor signaling,NF-κB activation, and tumor proliferation in chronic lymphocyticleukemia,” Blood 117(2):563-574 (2011); DAVIS et al., “Chronic activeB-cell-receptor signaling in diffuse large B-cell lymphoma,” Nature463:88-92 (2010); PIGHI et al., “Phospho-proteomic analysis of mantlecell lymphoma cells suggests a pro-survival role of B-cell receptorsignaling,” Cell Oncol. (Dordr) 34(2):141-153 (2011); RIZZATTI et al.,“Gene expression profiling of mantle cell lymphoma cells revealsaberrant expression of genes from the PI3K-AKT, WNT and TGFβ signalingpathways,” Brit. J. Haematol. 130:516-526 (2005); MARTINEZ et al., “TheMolecular Signature of Mantle Cell Lymphoma Reveals Multiple SignalsFavoring Cell Survival,” Cancer Res. 63:8226-8232 (2003). Interactionsbetween malignant B-cells and supporting cells (e.g., stromal cells,nurse-like cells) in the tumor microenvironment are important for tumorcell survival, proliferation, homing, and tissue retention. See, e.g.,BURGER, “Inhibiting B-Cell Receptor Signaling Pathways in ChronicLymphocytic Leukemia,” Curr. Mematol. Malig. Rep. 7:26-33 (2012);HERISHANU et al., “The lymph node microenvironment promotes B-cellreceptor signaling, NF-κB activation, and tumor proliferation in chroniclymphocytic leukemia,” Blood 117(2):563-574 (2011); KURTOVA et al.,“Diverse marrow stromal cells protect CLL cells from spontaneous anddrug-induced apoptosis: development of a reliable and reproduciblesystem to assess stromal cell adhesion-mediated drug resistance,” Blood114(20): 4441-4450 (2009); BURGER et al., “High-level expression of theT-cell chemokines CCL3 and CCL4 by chronic lymphocytic leukemia B cellsin nurselike cell cocultures and after BCR stimulation,” Blood 113(13)3050-3058 (2009); QUIROGA et al., “B-cell antigen receptor signalingenhances chronic lymphocytic leukemia cell migration and survival:specific targeting with a novel spleen tyrosine kinase inhibitor, R406,”Blood 114(5):1029-1037 (2009) Inhibiting PI3K-δ,γ with an inhibitor incertain malignant B-cells can block the BCR-mediated intracellularsurvival signaling as well as key interactions with theirmicroenvironment that are critical for their growth.

PI3K-δ and PI3K-γ also play a direct role in the survival andproliferation of certain T-cell malignancies. See, e.g., SUBRAMANIAM etal., “Targeting Nonclassical Oncogenes for Therapy in T-ALL,” CancerCell 21:459-472 (2012). Aberrant PI3K-δ and PI3K-γ activity provides thesignals necessary for the development and growth of certain T-cellmalignancies. While BTK is expressed in B-cells, it is not expressed inT-cells, and therefore BTK is not a viable target for the treatment ofT-cell malignancies. See, e.g., NISITANI et al., “Posttranscriptionalregulation of Bruton's tyrosine kinase expression in antigenreceptor-stimulated splenic B cells,” PNAS 97(6):2737-2742 (2000); DEWEERS et al., “The Bruton's tyrosine kinase gene is expressed throughoutB cell differentiation, from early precursor B cell stages precedingimmunoglobulin gene rearrangement up to mature B cell stages,” Eur. JImmunol. 23:3109-3114 (1993); SMITH et al., “Expression of Bruton'sAgammaglobulinemia Tyrosine Kinase Gene, BTK, Is SelectivelyDown-Regulated in T Lymphocytes and Plasma Cells,” J. Immunol.152:557-565 (1994). PI3K-δ and/or γ inhibitors may have uniquetherapeutic potential in T-cell malignancies.

In neutrophils, PI3K-δ, along with PI3K-γ, contribute to the responsesto immune complexes, FCγRII signaling, including migration andneutrophil respiratory burst. Human neutrophils undergo rapid inductionof PIP3 in response to formyl peptide receptor (FMLP) or complementcomponent C5a (C5a) in a PI3K-γ dependent manner, followed by a longerPIP3 production period that is PI3K-δ dependent, and is essential forrespiratory burst. The response to immune complexes is contributed byPI3K-δ, PI3K-γ, and PI3K-β, and is an important mediator of tissuedamage in models of autoimmune disease (Randis T M et al. (2008) Eur JImmunol. 38(5):1215-24; Pinho V, (2007) J Immunol. 179(11):7891-8; SadhuC. et al. (2003) J Immunol. 170(5):2647-54; Condliffe A M et al. (2005)Blood 106(4):1432-40). It has been reported that in certain autoimmunediseases, preferential activation of PI3K-β may be involved (Kulkarni etal., Immunology (2011) 4(168) ra23: 1-11). It was also reported thatPI3K-β-deficient mice were highly protected in an FcγR-dependent modelof autoantibody-induced skin blistering and partially protected in anFcγR-dependent model of inflammatory arthritis, whereas combineddeficiency of PI3K-β and PI3K-δ resulted in near complete protection ininflammatory arthritis (Id.).

In macrophages collected from patients with chronic obstructivepulmonary disease (COPD), glucocorticoid responsiveness can be restoredby treatment of the cells with inhibitors of PI3K-δ. Macrophages alsorely on PI3K-δ and PI3K-γ for responses to immune complexes through thearthus reaction (FCγR and C5a signaling) (Randis T M, et al. (2008) EurJ Immunol. 38(5):1215-24; Marwick J A et al. (2009) Am J Respir CritCare Med. 179(7):542-8; Konrad S, et al. (2008) J Biol Chem.283(48):33296-303).

Theophylline increases histone deacetylase-2 and corticosteroidsensitivity in vitro and in smoking mice in vivo by inhibiting PI3kinase (e.g., PI3K-delta). PI3K is activated in COPD lungs and certainPI3K inhibitors have been shown to mimic the effects of theophylline inreversing corticosteroid resistance. Yasuo, T., et al., Am J Respir CritCare Med 2010; 182:897-904. While not wishing to be bound by theory, arationale for the use of PI3K inhibitors (e.g., compounds providedherein) to treat COPD is that a PI3K inhibitor can increase thecorticosteroid sensitivity in a subject.

In mast cells, stem cell factor-(SCF) and IL3-dependent proliferation,differentiation and function are PI3K-δ dependent, as is chemotaxis. Theallergen/IgE crosslinking of FCγR1 resulting in cytokine release anddegranulation of the mast cells is severely inhibited by treatment withPI3K-δ inhibitors, suggesting a role for PI3K-δ in allergic disease (AliK et al. (2004) Nature 431(7011):1007-11; Lee K S, et al. (2006) FASEBJ. 20(3):455-65; Kim M S, et al. (2008) Trends Immunol. 29(10):493-501).

Natural killer (NK) cells are dependent on both PI3K-δ and PI3K-γ forefficient migration towards chemokines including CXCL10, CCL3, S1P andCXCL12, or in response to LPS in the peritoneum (Guo H, et al. (2008) JExp Med. 205(10):2419-35; Tassi I, et al. (2007) Immunity 27(2):214-27;Saudemont A, (2009) Proc Natl Acad Sci USA. 106(14):5795-800; Kim N, etal. (2007) Blood 110(9):3202-8).

The roles of PI3K-δ and PI3K-γ in the differentiation, maintenance, andactivation of immune cells support a role for these enzymes ininflammatory disorders ranging from autoimmune diseases (e.g.,rheumatoid arthritis, multiple sclerosis) to allergic inflammatorydisorders, such as asthma, and inflammatory respiratory disease, such asCOPD. Extensive evidence is available in experimental animal models, orcan be evaluated using art-recognized animal models. In an embodiment,described herein is a method of treating inflammatory disorders rangingfrom autoimmune diseases (e.g., rheumatoid arthritis, multiplesclerosis) to allergic inflammatory disorders, such as asthma and COPDusing a compound described herein.

For example, inhibitors of PI3K-δ and/or -γ have been shown to haveanti-inflammatory activity in several autoimmune animal models forrheumatoid arthritis (Williams, O. et al. (2010) Chem Biol,17(2):123-34; WO 2009/088986; WO2009/088880; WO 2011/008302; eachincorporated herein by reference). PI3K-δ is expressed in the RAsynovial tissue (especially in the synovial lining which containsfibroblast-like synoviocytes (FLS), and selective PI3K-δ inhibitors havebeen shown to be effective in inhibiting synoviocyte growth and survival(Bartok et al. (2010) Arthritis Rheum 62 Suppl 10:362). Several PI3K-δand -γ inhibitors have been shown to ameliorate arthritic symptoms(e.g., swelling of joints, reduction of serum-induced collagen levels,reduction of joint pathology and/or inflammation), in art-recognizedmodels for RA, such as collagen-induced arthritis and adjuvant inducedarthritis (WO 2009/088986; WO2009/088880; WO 2011/008302; eachincorporated herein by reference).

The role of PI3K-δ has also been shown in models of T-cell dependentresponse, including the DTH model. In the murine experimental autoimmuneencephalomyelitis (EAE) model of multiple sclerosis, the PI3K-γ/δ-doublemutant mice are resistant. PI3K-δ inhibitors have also been shown toblock EAE disease induction and development of TH-17 cells both in vitroand in vivo (Haylock-Jacobs, S. et al. (2011) J. Autoimmunity36(3-4):278-87).

Systemic lupus erythematosus (SLE) is a complex disease that atdifferent stages requires memory T-cells, B-cell polyclonal expansionand differentiation into plasma cells, and the innate immune response toendogenous damage associated molecular pattern molecules (DAMPS), andthe inflammatory responses to immune complexes through the complementsystem as well as the F_(C) receptors. The role of PI3K-δ and PI3K-γtogether in these pathways and cell types suggest that blockade with aninhibitor would be effective in these diseases. A role for PI3K in lupusis also predicted by two genetic models of lupus. The deletion ofphosphatase and tensin homolog (PTEN) leads to a lupus-like phenotype,as does a transgenic activation of Class 1A PI3Ks, which includesPI3K-δ. The deletion of PI3K-γ in the transgenically activated class 1Alupus model is protective, and treatment with a PI3K-γ selectiveinhibitor in the murine MLR//lpr model of lupus improves symptoms(Barber, D F et al. (2006) J. Immunol. 176(1): 589-93).

In allergic disease, PI3K-δ has been shown by genetic models and byinhibitor treatment to be essential for mast-cell activation in apassive cutaneous anaphalaxis assay (Ali K et al. (2008) J Immunol.180(4):2538-44; Ali K, (2004) Nature 431(7011):1007-11). In a pulmonarymeasure of response to immune complexes (Arthus reaction) a PI3K-δknockout is resistant, showing a defect in macrophage activation and C5aproduction. Knockout studies and studies with inhibitors for both PI3K-δand PI3K-γ support a role for both of these enzymes in the ovalbumininduced allergic airway inflammation and hyper-responsiveness model (LeeK S et al. (2006) FASEB J. 20(3):455-65). Reductions of infiltration ofeosinophils, neutrophils, and lymphocytes as well as TH2 cytokines (IL4,IL5, and IL13) were seen with both PI3K-δ specific and dual PI3K-δ andPI3K-γ inhibitors in the Ova induced asthma model (Lee K S et al. (2006)J Allergy Clin Immunol 118(2):403-9).

PI3K-δ and PI3K-γ inhibition can be used in treating COPD. In the smokedmouse model of COPD, the PI3K-δ knockout does not develop smoke inducedglucocorticoid resistance, while wild-type and PI3K-γ knockout mice do.An inhaled formulation of dual PI3K-δ and PI3K-γ inhibitor blockedinflammation in a LPS or smoke COPD models as measured by neutrophiliaand glucocorticoid resistance (Doukas J, et al. (2009) J Pharmacol ExpTher. 328(3):758-65).

PI3K-δ and/or PI3K-γ Isoforms in Certain Cancers

Class I PI3Ks, particularly PI3K-δ and PI3K-γ isoforms, are alsoassociated with cancers (reviewed, e.g., in Vogt, P K et al. (2010) CurrTop Microbiol Immunol. 347:79-104; Fresno Vara, J A et al. (2004) CancerTreat Rev. 30(2):193-204; Zhao, L and Vogt, PK. (2008) Oncogene27(41):5486-96) Inhibitors of PI3K, e.g., PI3K-δ and/or PI3K-γ, havebeen shown to have anti-cancer activity (e.g., Courtney, K D et al.(2010) J Clin Oncol. 28(6):1075-1083); Markman, B et al. (2010) AnnOncol. 21(4):683-91; Kong, D and Yamori, T (2009) Curr Med Chem.16(22):2839-54; Jimeno, A et al. (2009)J Clin Oncol. 27:156s (suppl;abstr 3542); Flinn, I W et al. (2009) J Clin Oncol. 27:156s (suppl;abstr 3543); Shapiro, G et al. (2009) J Clin Oncol. 27:146s (suppl;abstr 3500); Wagner, A J et al. (2009) J Clin Oncol. 27:146s (suppl;abstr 3501); Vogt, P K et al. (2006) Virology 344(1):131-8; Ward, S etal. (2003) Chem Biol. 10(3):207-13; WO 2011/041399; US 2010/0029693; US2010/0305096; US 2010/0305084; each incorporated herein by reference).

In one embodiment, described herein is a method of treating cancer. Inone embodiment, provided herein is a method of treating a hematologicalcancer comprising administering a pharmaceutically effective amount of acompound provided herein to a subject in need thereof. In oneembodiment, provided herein is a method of treating a solid tumorcomprising administering a pharmaceutically effective amount of acompound provided herein to a subject in need thereof. Types of cancerthat can be treated with an inhibitor of PI3K (e.g., Compound 1)include, e.g., leukemia, chronic lymphocytic leukemia, acute myeloidleukemia, chronic myeloid leukemia (e.g., Salmena, L et al. (2008) Cell133:403-414; Chapuis, N et al. (2010) Clin Cancer Res. 16(22):5424-35;Khwaja, A (2010) Curr Top Microbiol Immunol. 347:169-88); lymphoma,e.g., non-Hodgkin's lymphoma (e.g., Salmena, L et al. (2008) Cell133:403-414); lung cancer, e.g., non-small cell lung cancer, small celllung cancer (e.g., Herrera, V A et al. (2011) Anticancer Res.31(3):849-54); melanoma (e.g., Haluska, F et al. (2007) Semin Oncol.34(6):546-54); prostate cancer (e.g., Sarker, D et al. (2009) ClinCancer Res. 15(15):4799-805); glioblastoma (e.g., Chen, J S et al.(2008) Mol Cancer Ther. 7:841-850); endometrial cancer (e.g., Bansal, Net al. (2009) Cancer Control. 16(1):8-13); pancreatic cancer (e.g.,Furukawa, T (2008) J Gastroenterol. 43(12):905-11); renal cell carcinoma(e.g., Porta, C and Figlin, R A (2009) J Urol. 182(6):2569-77);colorectal cancer (e.g., Saif, M W and Chu, E (2010) Cancer J.16(3):196-201); breast cancer (e.g., Torbett, N E et al. (2008) BiochemJ. 415:97-100); thyroid cancer (e.g., Brzezianska, E andPastuszak-Lewandoska, D (2011) Front Biosci. 16:422-39); and ovariancancer (e.g., Mazzoletti, M and Broggini, M (2010) Curr Med Chem.17(36):4433-47).

Numerous publications support a role of PI3K-δ and PI3K-γ in treatinghematological cancers. PI3K-δ and PI3K-γ are highly expressed in theheme compartment, and solid tumors, including prostate, breast andglioblastomas (Chen J. S. et al. (2008) Mol Cancer Ther. 7(4):841-50;Ikeda H. et al. (2010) Blood 116(9): 1460-8).

In hematological cancers including acute myeloid leukemia (AML),multiple myeloma (MM), and chronic lymphocytic leukemia (CLL),overexpression and constitutive activation of PI3K-δ supports the modelthat PI3K-δ inhibition would be therapeutic Billottet C, et al. (2006)Oncogene 25(50):6648-59; Billottet C, et al. (2009) Cancer Res.69(3):1027-36; Meadows, S A, 52^(nd) Annual ASH Meeting and Exposition;2010 Dec. 4-7; Orlando, Fla.; Ikeda H, et al. (2010) Blood116(9):1460-8; Herman S E et al. (2010) Blood 116(12):2078-88; Herman SE et al. (2011). Blood 117(16):4323-7.

In one embodiment, described herein is a method of treatinghematological cancers including, but not limited to acute myeloidleukemia (AML), multiple myeloma (MM), and chronic lymphocytic leukemia(CLL).

A PI3K-δ inhibitor (CAL-101) has been evaluated in a phase 1 trial inpatients with hematological malignancies, and showed activity in CLL inpatients with poor prognostic characteristics. In CLL, inhibition ofPI3K-δ not only affects tumor cells directly, but it also affects theability of the tumor cells to interact with their microenvironment. Thismicroenvironment includes contact with and factors from stromal cells,T-cells, nurse like cells, as well as other tumor cells. CAL-101suppresses the expression of stromal and T-cell derived factorsincluding CCL3, CCL4, and CXCL13, as well as the CLL tumor cells'ability to respond to these factors. CAL-101 treatment in CLL patientsinduces rapid lymph node reduction and redistribution of lymphocytesinto the circulation, and affects tonic survival signals through theBCR, leading to reduced cell viability, and an increase in apoptosis.Single agent CAL-101 treatment was also active in mantle cell lymphomaand refractory non-Hodgkin's lymphoma (Furman, R R, et al. 52^(nd)Annual ASH Meeting and Exposition; 2010 Dec. 4-7; Orlando, Fla.;Hoellenriegel, J, et al. 52^(nd) Annual ASH Meeting and Exposition; 2010Dec. 4-7; Orlando, Fla.; Webb, H K, et al. 52^(nd) Annual ASH Meetingand Exposition; 2010 Dec. 4-7; Orlando, Fla.; Meadows, et al. 52^(nd)Annual ASH Meeting and Exposition; 2010 Dec. 4-7; Orlando, Fla.; Kahl,B, et al. 52^(nd) Annual ASH Meeting and Exposition; 2010 Dec. 4-7;Orlando, Fla.; Lannutti B J, et al. (2011) Blood 117(2):591-4).

PI3K-δ inhibitors have shown activity against PI3K-δ positive gliomas invitro (Kashishian A, et al. Poster presented at: The AmericanAssociation of Cancer Research 102^(nd) Annual Meeting; 2011 Apr. 2-6;Orlando, Fla.). In this subset of tumors, treatment with the PI3K-δinhibitor either alone or in combination with a cytotoxic agent can beeffective.

Another mechanism for PI3K-δ inhibitors to have an effect in solidtumors involves the tumor cells' interaction with theirmicro-environment. PI3K-δ, PI3K-γ, and PI3K-β are expressed in theimmune cells that infiltrate tumors, including tumor infiltratinglymphocytes, macrophages, and neutrophils. PI3K-δ inhibitors can modifythe function of these tumor-associated immune cells and how they respondto signals from the stroma, the tumor, and each other, and in this wayaffect tumor cells and metastasis (Hoellenriegel, J, et al. 52^(nd)Annual ASH Meeting and Exposition; 2010 Dec. 4-7; Orlando, Fla.).

PI3K-δ is also expressed in endothelial cells. It has been shown thattumors in mice treated with PI3K-δ selective inhibitors are killed morereadily by radiation therapy. In this same study, capillary networkformation is impaired by the PI3K inhibitor, and it is postulated thatthis defect contributes to the greater killing with radiation. PI3K-δinhibitors can affect the way in which tumors interact with theirmicroenvironment, including stromal cells, immune cells, and endothelialcells and be therapeutic either on its own or in conjunction withanother therapy (Meadows, S A, et al. Paper presented at: 52^(nd) AnnualASH Meeting and Exposition; 2010 Dec. 4-7; Orlando, Fla.; Geng L, et al.(2004) Cancer Res. 64(14):4893-9).

Accordingly, provided herein is a method of treating or preventing acancer or disease (including but not limited to a hematologicmalignancy, or a specific type or sub-type of cancer or disease, such asa specific type or sub-type of hematologic malignancy), with a PI3K-γselective inhibitor, wherein the adverse effects associated withadministration of inhibitors for other isoform(s) of PI3K (e.g., PI3K-αand/or PI3K-β) are reduced. In one embodiment, provided herein is amethod of treating or preventing the cancer or disease, with a PI3K-γselective inhibitor, at a lower (e.g., by about 10%, by about 20%, byabout 30%, by about 40%, by about 50%, by about 60%, by about 70%, or byabout 80%) dose as compared to treatment with a PI3K-γ non-selective orless selective PI3K-γ inhibitor (e.g., a PI3Kpan inhibitors, e.g.,inhibiting PI3K-α, β, δ, and γ).

The role of PI3K-γ pathway in promoting myeloid cell trafficking totumors and the role of blockade of p100γ in suppression of tumorinflammation and growth in breast cancer, pancreatic cancer, and lungcancer are reported, for example, in Schmid et al. (2011) Cancer Cell19, 715-727, the entirety of which is incorporated herein by reference.In one embodiment, provided herein is a method of treating or preventingpancreatic cancer with a PI3K inhibitor.

In another embodiment, provided herein is a method of treating orpreventing breast cancer with a PI3K inhibitor. In yet anotherembodiment, provided herein is a method of treating or preventing lungcancer with a PI3K inhibitor. In one embodiment, the PI3K inhibitor is aPI3K-γ inhibitor, selective or non-selective over one or more other PI3Kisoform(s). In one embodiment, the PI3K inhibitor is a PI3K-γ selectiveinhibitor.

While not wishing to be bound by theory, it is believed that tumorgrowth is influenced by two classes of immune cells in the tumormicroenvironment: effector cells which include cytotoxic cells and M1macrophages, and which have anti-tumor activity, and suppressor cells,which include M2 macrophages, MDSC (myeloid derived suppressor cell),Tregs (regulatory T cell), and regulatory dendritic cells, which havepro-tumor activity because they inhibit the effector cells. An abundanceof suppressor cells can lead to tumor immune tolerance, and enhancementof tumor growth.

Certain of these cell types are briefly described. M1 denotes apro-inflammatory (anti-tumor) phenotype of a MDSC or TAM. M2 denotes ananti-inflammatory (pro-tumor) phenotype of a MDSC or TAM.

PI3K-γ is not expressed in at least some cancer cell types. Schmid etal., 2011, Cancer Cell 19. Accordingly, in some embodiments, the PI3K-γinhibitor reduces cancer cell growth without having a substantial directeffect on the cancer cell itself. For instance, in some embodiments, thePI3K-γ inhibitor inhibits cancer cell growth through changes in thetumor microenvironment, e.g., the immune cells in close proximity to thecancer cells.

Evidence in the literature supports the idea that a PI3K-γ inhibitor canreduce tumor associated myeloid cells. For instance, in PI3K-γ-deficientmice, tumor-associated myeloid cells are reduced. Schmid et al., 2011,Cancer Cell 19. Together, these data indicate that a large class ofPI3K-γ inhibitors should reduce tumor associated myeloid cells, therebyincreasing the immune response against cancer cells, and treating thecancer. While not wishing to be bound by theory, a PI3K-γ may operatethrough the following mechanism. PI3K-γ signaling may tilt the balanceof immune cells towards pro-tumor M2 cells and away from anti-tumor M1cells, by inducing expression of immunosuppressive, wound healing genessuch as Arginase 1, TGFbeta1, PDGFBB, MMP9, and MMP13, and suppressingpro-inflammatory factors such as IL12, iNos, and interferon gamma.Blocking PI3K-γ signaling with an inhibitor tilts the balance towardsanti-tumor M1 cells by stimulating a T cell activating gene expressionprogram. Kaneda et al. PI3-kinase gamma controls the macrophage M1-M2switch, thereby promoting tumor immunosuppression and progression.[abstract]. In: Proceedings of the 105th Annual Meeting of the AmericanAssociation for Cancer Research; 2014 Apr. 5-9; San Diego, Calif.Philadelphia (Pa.): AACR; Cancer Res 2014; 74(19 Suppl):Abstract nr3650. doi:10.1158/1538-7445.AM2014-3650.

In some embodiments, a PI3K-γ inhibitor provided herein is administeredto a patient in order to block a homeostatic down-regulation of T cellresponse. While not wishing to be bound by theory, this may allow thebody to raise an effective immune response against the cancer cell.Exemplary agents of this type include immune checkpoint therapeutics,e.g., agents that act on CTLA-4, PD-1, or PD-L1, e.g., antibodies thatbind to CTLA-4, PD-1, or PD-L1. Immune checkpoint therapeutics aredescribed in more detail below.

In some embodiments, a PI3K-γ inhibitor provided herein is administeredto a patient in order to eliminate immunosuppressive cells in the tumormicroenvironment. The immunosuppressive cell may be, e.g., a Tregulatory cell (e.g., a cell that secretes mediators that induce CD8+cytotoxic T cell death); a Tumor-associated macrophage (TAM; e.g., anM2(pro-tumor) TAMS that blocks T cell activity and promotes angiogenesis);or a myeloid-derived suppressor cell (MDSC; e.g., a cell that secretesmediators that inhibit T cell differentiation and proliferation).

In some embodiments, a compound provided herein is administered to apatient in order to reduce the migration or differentiation of a tumorassociated myeloid cell. In some embodiments, the compound is a compoundthat shows single agent activity in a syngeneic model system. In someembodiments, the compound is administered in combination with a secondtherapeutic, as discussed herein. In some embodiments, theadministration results in a reduction in the level of MDSCs in the tumormicroenvironment; the level of M2 TAMS in the tumor microenvironment;the level of T-regulatory cells in the tumor microenvironment, or anycombination thereof. In some embodiments, the administration results inan unchanged or increased level of T-effector cells in the tumormicroenvironment. In embodiments, the administration results in anincrease in an immune response to the tumor, e.g., an increase in thelevels or tumor-attacking activity of cytotoxic T cells, M1 inflammatoryTAMs, or a combination thereof.

In some embodiments, an MDSC has one or more of the followingproperties: suppressing anti-tumor immune attack; inducingvascularization of the tumor; inducing ECM breakdown, e.g., which maycontribute to metastasis; and supporting tumor growth. Accordingly, insome embodiments, administration of a PI3K-γ inhibitor described hereininhibits one or more of these functions in an MDSC.

TAMs (tumor-associated macrophages) can also have one or more of thefollowing properties: suppressing anti-tumor immune attack; inducingvascularization of the tumor; inducing ECM breakdown, e.g., which maycontribute to metastasis; and supporting tumor growth. Accordingly, insome embodiments, administration of a PI3K-γ inhibitor as describedherein inhibits one or more of these functions in a TAM.

In embodiments, a PI3K-γ inhibitor is administered to a patient who hasreceived chemotherapy and/or radiation therapy. While not wishing to bebound by theory, in some embodiments, chemotherapy or radiation therapyresults in a wound healing response that leads to repopulation of thecancer site, e.g., tumor, with TAMs and MDSCs. Administering the PI3K-γinhibitor, in some embodiments, reduces the levels of TAMs and MDSCs inthe microenvironment, decreasing their support for tumor cell growthand/or allowing the immune system to attack the cancer cells. See ClaireE. Lewis, “Imaging immune cell infiltrating tumors in zebrafish”, AACRAnnual Meeting (Apr. 5, 2014).

While not wishing to be bound by theory, a rationale for the use of aPI3K-gamma inhibitor as adjunct therapy to radiation is to prevent theaccumulation of tumor supporting myeloid cells into the radiated tumor,thus impairing tumor regrowth following radiation therapy. This issupported by work of Kioi et al. (2010) Clin Invest. 120(3):694-705showing that an inhibitor of myeloid cell migration into post-irradiatedtumors (e.g., AMD3100) blocked tumor vasculogenesis and tumor regrowth.

In certain embodiments, provided herein is a method of treating adisorder or disease provided herein, comprising administering a compoundprovided herein, e.g., a PI3K γ selective inhibitor, a PI3K δ selectiveinhibitor, or a PI3K γ/δ dual inhibitor. Without being limited by aparticular theory, in some embodiments, selectively inhibiting PI3K-γisoform can provide a treatment regimen where adverse effects associatedwith administration of a non-selective PI3K inhibitor are minimized orreduced. Without being limited by a particular theory, in someembodiments, selectively inhibiting PI3K-δ isoform can provide atreatment regimen where adverse effects associated with administrationof a non-selective PI3K inhibitor are minimized or reduced. Withoutbeing limited by a particular theory, in some embodiments, selectivelyinhibiting PI3K-δ and γ isoform can provide a treatment regimen whereadverse effects associated with administration of a non-selective PI3Kinhibitor are minimized or reduced. Without being limited by aparticular theory, it is believed that the adverse effects can bereduced by avoiding the inhibition of other isoforms (e.g., α or β) ofPI3K.

In one embodiment, the adverse effect is hyperglycemia. In anotherembodiment, the adverse effect is rash. In another embodiment, theadverse effect is impaired male fertility that may result frominhibition of β isoform of PI3K (see, e.g., Ciraolo et al., MolecularBiology of the Cell, 21: 704-711 (2010)). In another embodiment, theadverse effect is testicular toxicity that may result from inhibition ofPI3K-β (see, e.g., Wisler et al., Amgen SOT, Abstract ID #2334 (2012)).In another embodiment, the adverse effect is embryonic lethality (see,e.g., Bi et al., J Biol Chem, 274: 10963-10968 (1999)). In anotherembodiment, the adverse effect is defective platelet aggregation (see,e.g., Kulkarni et al., Science, 287: 1049-1053 (2000)). In anotherembodiment, the adverse effect is functionally defective neutrophil(id).

In certain embodiments, provided herein is a method of treating orpreventing cancer (e.g., colon cancer, melanoma, bladder cancer, renalcancer, breast, lung cancer, glioblastoma, solid tumors, and a cancer ofhematopoietic origin (e.g., DLBCL, CLL, Hodgkin lymphoma, non-Hodgkinlymphomas)) comprising administering to the subject a PI3K inhibitor(e.g., a PI3K-γ inhibitor, e.g., Compound 1).

Without being bound by a particular theory, a rationale for the use of aPI3K inhibitor to treat or prevent cancer is that cells derived fromtumors (e.g., from CT26 mouse tumors) can suppress anti-tumor immunecell function, including T-cell proliferation, as shown in the examplesprovide herein, and treatment with a compound provided herein canrelease the suppression. The tumor microenvironment can inhibit theactivation and proliferation of immune effector cells due to thepresence of suppressive myeloid cells (e.g., myeloid derived suppressorcells or MDSC and M2 macrophages). Compounds provided herein can affectthe number and activity M2 macrophages in a tumor microenvironment,e.g., reduce or inhibit the level of M2, pro-tumor macrophages. Thereduction or inhibition of M2 macrophages, which produceanti-inflammatory cytokines and other factors, would lead to increasedanti-tumor immunity, including T cell proliferation. Therefore, acompound provided herein can treat or prevent cancer such as coloncancer, melanoma, bladder cancer, renal cancer, breast, lung cancer,glioblastoma, solid tumors, and a cancer of hematopoietic origin (e.g.,lymphoma, DLBCL, CLL, Hodgkin disease, non-Hodgkin lymphomas). Further,it has also been shown in the examples provided herein that anti-PDL1can also release suppression of T cell proliferation by blocking theinteraction between PD1 on T cells and PDL1 on tumor cells andregulatory cells. The cytotoxic T cells that are induced to proliferateand survive by both anti PDL-1 and compound 1 are hypothesized to slowtumor growth. Compounds provided herein can relieve immunosuppressionwhich can lead to T cells proliferation and activation. Compoundsprovided herein can treat or prevent cancer by inducing T cell mediatedimmunity. In one embodiment, the compound provided herein can decreasetumor volume. In one embodiment, a combination of a PI3K inhibitor suchas a compound provided herein and anti-PDL1 would be effective intreating or preventing cancer by inducing T cell mediated tumorimmunity. In some embodiments, the effect of a compound provided hereinon T-cell function can be assessed by analyzing the pro-inflammatorycytokine levels in tumor tissues and serum, e.g., a MSD pro-inflammatorypanel. In another embodiment, the pro-inflammatory cytokines areselected from IFN-γ, IL-1β, IL-10, IL-12 p70, IL-2, IL-4, IL-5, IL-6,KC/GRO, and TNF-α. In one embodiment, the effect of a compound providedherein on T cell function can be assessed by analyzing the IFN-γ level.For example, tumor tissues and serum treated with a compound providedherein, e.g., Compound 1, can be assessed by analyzing the IFN-γ level.

Treatment of Neuropsychiatric Disorders

In other embodiments, inhibition of PI3K (such as PI3K-δ and/or PI3K-γ)can be used to treat a neuropsychiatric disorder, e.g., an autoimmunebrain disorder. Infectious and immune factors have been implicated inthe pathogenesis of several neuropsychiatric disorders, including, butnot limited to, Sydenham's chorea (SC) (Garvey, M. A. et al. (2005) J.Child Neurol. 20:424-429), Tourette's syndrome (TS), obsessivecompulsive disorder (OCD) (Asbahr, F. R. et al. (1998) Am. J. Psychiatry155:1122-1124), attention deficit/hyperactivity disorder (AD/HD)(Hirschtritt, M. E. et al. (2008) Child Neuropsychol. 1:1-16; Peterson,B. S. et al. (2000) Arch. Gen. Psychiatry 57:364-372), anorexia nervosa(Sokol, M. S. (2000) J. Child Adolesc. Psychopharmacol. 10:133-145;Sokol, M. S. et al. (2002) Am. J. Psychiatry 159:1430-1432), depression(Leslie, D. L. et al. (2008) J. Am. Acad. Child Adolesc. Psychiatry47:1166-1172), and autism spectrum disorders (ASD) (Hollander, E. et al.(1999) Am. J. Psychiatry 156:317-320; Margutti, P. et al. (2006) Curr.Neurovasc. Res. 3:149-157). A subset of childhood obsessive compulsivedisorders and tic disorders has been grouped as Pediatric AutoimmuneNeuropsychiatric Disorders Associated with Streptococci (PANDAS). PANDASdisorders provide an example of disorders where the onset andexacerbation of neuropsychiatric symptoms is preceded by a streptococcalinfection (Kurlan, R., Kaplan, E. L. (2004) Pediatrics 113:883-886;Garvey, M. A. et al. (1998) J. Clin. Neurol. 13:413-423). Many of thePANDAS disorders share a common mechanism of action resulting fromantibody responses against streptococcal associated epitopes, such asGlcNAc, which produces neurological effects (Kirvan. C. A. et al. (2006)J. Neuroimmunol. 179:173-179). Autoantibodies recognizing centralnervous system (CNS) epitopes are also found in sera of most PANDASsubjects (Yaddanapudi, K. et al. (2010) Mol. Psychiatry 15:712-726).Thus, several neuropsychiatric disorders have been associated withimmune and autoimmune components, making them suitable for therapiesthat include PI3K-δ and/or PI3K-γ inhibition.

In certain embodiments, a method of treating (e.g., reducing orameliorating one or more symptoms of) a neuropsychiatric disorder,(e.g., an autoimmune brain disorder), using a PI3K-δ and/or PI3K-γinhibitor is described, alone or in combination therapy. For example,one or more PI3K-δ and/or PI3K-γ inhibitors described herein can be usedalone or in combination with any suitable therapeutic agent and/ormodalities, e.g., dietary supplement, for treatment of neuropsychiatricdisorders. Exemplary neuropsychiatric disorders that can be treated withthe PI3K-δ and/or PI3K-γ inhibitors described herein include, but arenot limited to, PANDAS disorders, Sydenham's chorea, Tourette'ssyndrome, obsessive compulsive disorder, attention deficit/hyperactivitydisorder, anorexia nervosa, depression, and autism spectrum disorders.Pervasive Developmental Disorder (PDD) is an exemplary class of autismspectrum disorders that includes Autistic Disorder, Asperger's Disorder,Childhood Disintegrative Disorder (CDD), Rett's Disorder and PDD-NotOtherwise Specified (PDD-NOS). Animal models for evaluating the activityof the PI3K-δ and/or PI3K-γ inhibitor are known in the art. For example,a mouse model of PANDAS disorders is described in, e.g., Yaddanapudi, K.et al. (2010) supra; and Hoffman, K. I. et al. (2004) J. Neurosci.24:1780-1791.

In some embodiments, provided herein is a method for treating rheumatoidarthritis or asthma in a subject, or for reducing a rheumatoidarthritis-associated symptom or an asthma-associated symptom in asubject, comprising administering an effective amount of a PI3K-γinhibitor to a subject in need thereof, wherein one or more of theadverse effects associated with administration of inhibitors for one ormore other isoforms of PI3K are reduced. In one embodiment, the one ormore other isoforms of PI3K is PI3K-α, PI3K-β, and/or PI3K-δ. In oneembodiment, the one or more other isoforms of PI3K is PI3K-α and/orPI3K-β. In one embodiment, the method is for treating rheumatoidarthritis in a subject, or for reducing a rheumatoidarthritis-associated symptom in a subject. In another embodiment, themethod is for treating asthma in a subject, or for reducing anasthma-associated symptom in a subject.

In some embodiments, provided herein are methods of using a compoundprovided herein, or a pharmaceutically acceptable form (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein, to treat diseaseconditions, including, but not limited to, diseases associated withmalfunctioning of one or more types of PI3 kinase. In one embodiment, adetailed description of conditions and disorders mediated by p110δkinase activity is set forth in Sadu et al., WO 01/81346, which isincorporated herein by reference in its entirety for all purposes.

In some embodiments, the disclosure relates to a method of treating ahyperproliferative disorder in a subject that comprises administering tosaid subject a therapeutically effective amount of a compound providedherein, or a pharmaceutically acceptable form (e.g., pharmaceuticallyacceptable salts, hydrates, solvates, isomers, prodrugs, andisotopically labeled derivatives) thereof, or a pharmaceuticalcomposition as provided herein. In some embodiments, said method relatesto the treatment of cancer such as acute myeloid leukemia, thymus,brain, lung, squamous cell, skin, eye, retinoblastoma, intraocularmelanoma, oral cavity and oropharyngeal, bladder, gastric, stomach,pancreatic, bladder, breast, cervical, head, neck, renal, kidney, liver,ovarian, prostate, colorectal, esophageal, testicular, gynecological,thyroid, CNS, PNS, AIDS-related (e.g., Lymphoma and Kaposi's Sarcoma) orviral-induced cancer. In some embodiments, said method relates to thetreatment of a non-cancerous hyperproliferative disorder such as benignhyperplasia of the skin (e.g., psoriasis), restenosis, or prostate(e.g., benign prostatic hypertrophy (BPH)).

Treatment of Cancers

In certain embodiments, provided herein are methods of modulating tumormicroenvironment of cancer cells in a subject, comprising administeringto the subject a therapeutically effective amount of a compound providedherein (e.g., a selective PI3K-γ inhibitor, e.g., Compound 1), or apharmaceutically acceptable form thereof.

As used herein and unless otherwise specified, “tumor microenvironment”refers to the cellular and extracellular environment where the tumorsare located. This location can include surrounding blood vessels, immunecells, fibroblasts, secreted signaling molecules, and the extracellularmatrix. The tumor microenvironment includes non-neoplastic stromal andimmune cells that provide growth and survival support to the neoplastictumor.

As used herein and unless otherwise specified, “immunotherapy” refers totreatments that stimulate, enhance, or suppress the body's own immunesystem to fight a disease. Diseases that may be suitable forimmunotherapy treatment include, but are not limited to, cancer,inflammatory diseases, and infectious diseases. Immunotherapy includes avariety of treatments that work in different ways. For example, some areintended to boost the immune system defenses in a general way; othershelp train the immune system to recognize and attack cancer cellsspecifically. Cancer immunotherapies include, but are not limited to,cell-based therapies (also known as cancer vaccines), antibodytherapies, and cytokine therapies (e.g., interleukin-2 andinterferon-α).

Many cancers are known to be susceptible to the treatment of one or moreimmunotherapies, including treatment targeting the effector cells in thetumor microenvironment (e.g., immune checkpoint therapy such asPD-1/PD-L1 inhibitors and CTLA-4 inhibitors), treatment targetingsuppressor cells in the tumor microenvironment (e.g., CSF-1R inhibitors(affecting MDSC and TAM) and IDO/TDO inhibitors). Without being limitedby a particular theory, a compound provided may affect MDSC, TAM, andother components in the tumor microenvironment. The role of TAM in tumormicroenvironment is described, e.g., in Lewis and Pollard, Cancer Res.2006; 66: (2). Jan. 15, 2006.

In one embodiment, the number of one or more pro-tumor immune cells inthe tumor microenvironment is reduced, or the activity of one or morepro-tumor immune cells in the tumor microenvironment is reduced orinhibited, after administration of the compound.

In some embodiments, the pro-tumor immune cell is a T-cell, an M2macrophage, a stromal cell, a dendritic cell, an endothelial cell, or amyeloid cell. In one embodiment, the myeloid cell is a tumor associatedsuppressive myeloid cell. In one embodiment, the tumor associatedsuppressive myeloid cell is identified by (i) CD45+, CD11b+, Ly6C+ andLy6G+, (ii) CD45+, CD11b+, Ly6C− and Ly6G−, (iii) CD45+, CD11b+, Ly6C−and Ly6G+, or (iv) CD45+, CD11b+, Ly6C+ and Ly6G−. In one embodiment,the tumor associated suppressive myeloid cell is a tumor associatedmacrophage (TAM), a myeloid derived suppressor cell (MDSC), a monocyticimmature myeloid cell (iMc), or a granulocytic iMc/neutrophil. In oneembodiment, the TAM is identified by CD45+, CD11b+, Ly6C−, and Ly6G−. Inone embodiment, the myeloid derived suppressor cell (MDSC) is identifiedby CD45+, CD11b+, Ly6C− and Ly6G+. In one embodiment, the monocyticimmature myeloid cell (iMc) is identified by CD45+, CD11b+, Ly6C+ andLy6G−. In one embodiment, the granulocytic iMc/neutrophil is identifiedby CD45+, CD11b+, Ly6C+ and Ly6G+. See e.g., Coussens L M. et al.,Cancer Discov. 2011 June; 1(1):54-67.

In one embodiment, the activation of M2 macrophage in the tumormicroenvironment is reduced or inhibited after administration of thecompound. In one embodiment, the p-AKT level in the M2 macrophage isreduced after administration of the compound. In one embodiment, thenumber of M2 macrophage cells in the tumor microenvironment is reducedafter administration of the compound. In one embodiment, the migrationof M2 macrophage cells into the tumor microenvironment is reduced orinhibited after administration of the compound. In one embodiment, thedifferentiation of myeloid cells into M2 macrophage cells in the tumormicroenvironment is reduced or inhibited after administration of thecompound. In one embodiment, the differentiation into M2 macrophagecells is measured by Arginase-1 (ARG1) level or VEGF level, and the ARG1level or VEGF level is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, or 90% compared to a reference value.

In one embodiment, the number of myeloid-derived suppressor cells in thetumor microenvironment is reduced after administration of the compound.In one embodiment, the differentiation of bone marrow cells intomyeloid-derived suppressor cells is reduced or inhibited afteradministration of the compound. In one embodiment, the differentiationinto myeloid-derived suppressor cells is measured by Arginase-1 (ARG1)level, VEGF level, or iNOS level, and the ARG1 level, VEGF level, oriNOS level is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, or 90% compared to a reference value.

In one embodiment, the production of proangiogenic factor is reduced orinhibited after administration of the compound. In one embodiment, theproangiogenic factor is reduced or inhibited by reduction or inhibitionof macrophage or MDSC differentiation. In one embodiment, theproangiogenic factor is VEGF.

In one embodiment, the effect of the compounds provided herein (e.g.,Compound 1) on MDSC (e.g., human MDSC) function is measured byexpression of iNOS and arginase and production of ROS and IL-10,measured by the suppression function of the MDSC (e.g., in co-cultureassays with CD8+), measured by activation of pAKT in response to astimulant (e.g., CXCL12, IL-1b, TNF-α, or CSF1), or measured bytranswell chemotaxis assays (T cells and MDSC).

In one embodiment, the effect of the compounds provided herein (e.g.,Compound 1) on MDSC (e.g., murine MDSC) function and macrophageM2-polarization is measured by isolating myeloid cells from bone marrow,polarizing with IFNg or IL-4 and then testing for secretion of TNF-α,IL-12, ROS production in M1 and IL-10, IL-1b, or VEGF, or measured bymethods provided herein or elsewhere.

In one embodiment, the effect of the compounds provided herein (e.g.,Compound 1) on myeloid and CD8+ is measured by in vivo models (e.g.,MC38 and 4T1). In one embodiment, the effect is measured by TGI, MDSCand macrophage infiltrate, CD8+, and IFN-gamma production in CD8+.

In one embodiment, the effect of the compounds provided herein (e.g.,Compound 1) on myeloid and CD8+ is measured by QT-PCR or intracellularFACS of myeloid infiltrate. In one embodiment, the effect is measured byexpression of functional markers (e.g., iNOS, arginase, or IL-10).

In one embodiment, the number of one or more anti-tumor immune cells inthe tumor microenvironment is increased, or the activity of one or moreanti-tumor immune cells in the tumor microenvironment is increased,after administration of the compound.

In one embodiment, the cancer susceptible to the treatment of one ormore immunotherapies is a hematological cancer. In one embodiment, thehematological cancer is chronic lymphocytic leukemia (CLL). In oneembodiment, the tumor microenvironment is a CLL proliferation center. Inone embodiment, the hematological cancer is lymphoma.

In one embodiment, the cancer susceptible to the treatment of one ormore immunotherapies is a solid tumor. In one embodiment, the solidtumor is lung cancer, breast cancer, colon cancer, or glioblastoma. Inone embodiment, the cancer is selected from one or more of: a cancer ofthe pulmonary system, a brain cancer, a cancer of the gastrointestinaltract, a skin cancer, a genitourinary cancer, a pancreatic cancer, alung cancer, a medulloblastoma, a basal cell carcinoma, a glioma, abreast cancer, a prostate cancer, a testicular cancer, an esophagealcancer, a hepatocellular cancer, a gastric cancer, a gastrointestinalstromal tumor (GIST), a colon cancer, a colorectal cancer, an ovariancancer, a melanoma, a neuroectodermal tumor, head and neck cancer, asarcoma, a soft-tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma,a chondrosarcoma, an osteogenic sarcoma, a chordoma, an angiosarcoma, anendotheliosarcoma, a lymphangiosarcoma, a lymphangioendotheliosarcoma, asynovioma, a mesothelioma, a leiomyosarcoma, a cervical cancer, auterine cancer, an endometrial cancer, a carcinoma, a bladder carcinoma,an epithelial carcinoma, a squamous cell carcinoma, an adenocarcinoma, abronchogenic carcinoma, a renal cell carcinoma, a hepatoma, a bile ductcarcinoma, a neuroendocrine cancer, a carcinoid tumor, diffuse typegiant cell tumor, and glioblastoma. In one embodiment, the solid tumoris melanoma, bladder cancer, head and neck cancer, lung cancer (e.g.,non-small cell lung cancer), renal cell carcinoma, ovarian cancer,breast cancer (e.g., triple-negative breast cancer), colon cancer, orglioblastoma.

In one embodiment, the solid tumor is melanoma. In one embodiment, thesolid tumor is lung cancer. In one embodiment, the solid tumor isnon-small cell lung cancer. In one embodiment, the solid tumor is renalcell carcinoma. Melanoma, lung cancer (e.g., non-small cell lungcancer), and renal cell carcinoma are known to be sensitive toimmunotherapies. Data linking a poor prognosis to high TAM cell countshave been reported in breast, prostate, endometrial, bladder, kidney,esophageal, superficial, carcinoma, melanoma, and follicular lymphomacancers. See e.g., Lewis and Pollard, Cancer Res. 2006; 66: (2). Jan.15, 2006. One anti-PD-1 antibody drug, nivolumab, (Opdivo—Bristol MyersSquibb), produced complete or partial responses in non-small-cell lungcancer, melanoma, and renal-cell cancer, in a clinical trial with atotal of 296 patients.

In one embodiment, the solid tumor is head and neck cancer. Head andneck tumors tend to be highly immunogenic and have stronganti-PD-1/PD-L1 efficacy. In one embodiment, the solid tumor is bladdercancer. Bladder cancer also has strong anti-PD-1/PD-L1 efficacy. A highnumber of TAM cells has been associated with a poor prognosis andincreased tumor angiogenesis in bladder cancer.

In one embodiment, the solid tumor is breast cancer. In one embodiment,the breast cancer is triple-negative breast cancer. A high number of TAMcells has been associated with a poor prognosis of breast cancer. Seee.g., Lewis and Pollard, Cancer Res. 2006; 66: (2). Jan. 15, 2006. Inone embodiment, the solid tumor is ovarian cancer. In one embodiment,the solid tumor is colon cancer. Breast cancer, ovarian cancer, andcolon cancer are known to be sensitive to immunotherapies (e.g.,bevacizumab and trastuzumab) and can also have anti-PD-1/PD-L1 efficacy.

In one embodiment, the solid tumor is glioblastoma. In one embodiment,the solid tumor is glioblastoma multiforme. It has been reported thatPI3K-gamma expression is upregulated in brain microglia. Without beinglimited by a particular theory, PI3K-γ inhibitors provided herein (e.g.,Compound 1) may have P-glycoprotein inhibitory activity and thus cancross the blood brain barrier.

In one embodiment, the anti-tumor immune attack by effector T cells isincreased, vascularization of the tumor is reduced, extracellular matrix(ECM) breakdown is reduced, or tumor growth is decreased, compared to areference value, after administration of the compound.

In one embodiment, the tumor volume of the cancer is reduced afteradministration of the compound. In one embodiment, the tumor volume ofthe cancer is reduced by at least 10%, 20%, 30%, 50%, 60%, or 60%,compared to a reference value.

In one embodiment, the level of apoptosis of the cancer cells isincreased after administration of the compound. In one embodiment, thelevel of apoptosis of the cancer cells is increased by at least 10%,20%, 30%, 40%, or 50%, compared to a reference value.

In some embodiments, the subject is naive to immunotherapy treatment. Insome embodiments, the subject is naive to radiation therapy treatment.In some embodiments, the subject is naive to chemotherapy treatment.

In some embodiments, the subject has been pre-treated or previouslytreated with one or more immunotherapy treatments. In one embodiment,the subject is responsive to the pre-treatment or previous treatmentwith the immunotherapy. In one embodiment, the immunotherapy treatmentis a checkpoint treatment such as a PD-1 or PD-L1 inhibitor. In oneembodiment, the subject is a smoker. It has been reported that smokerpatients may respond better to immunotherapy (e.g., a PD-L1 inhibitorMPDL3280A) than non-smoker patients in a phase I clinical study forpatients with melanoma or cancers of the lung, kidney, colon, GI tract,or head and neck cancers.

In one embodiment, the cancer is melanoma, and the subject has beenpre-treated or previously treated with one or more immunotherapytreatments. In one embodiment, the subject has been pre-treated orpreviously treated with two or more immunotherapy treatments.

In one embodiment, the cancer is head and neck cancer, lung cancer(e.g., non-small cell lung cancer), renal cell carcinoma, or bladdercancer, and the subject has been pre-treated or previously treated withone immunotherapy treatment.

In one embodiment, the cancer is breast cancer (e.g., triple-negativebreast cancer), ovarian cancer, glioblastoma, or colon cancer, and thesubject is naive to immunotherapy treatment.

In one embodiment, provided herein is a method of treating, preventing,or managing melanoma in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., Compound 1), or a pharmaceutically acceptable form thereof,wherein the subject has been pre-treated or previously treated with oneor more immunotherapy treatments. In one embodiment, the immunotherapytreatment is ipilimumab (Yervoy), interleukin-2, vemurafenib,dabrafenib, or trametinib.

In one embodiment, provided herein is a method of treating, preventing,or managing lung cancer (e.g., non-small cell lung cancer) in a subject,comprising administering to the subject a therapeutically effectiveamount of a compound provided herein (e.g., Compound 1), or apharmaceutically acceptable form thereof, wherein the subject has beenpre-treated or previously treated with one or more immunotherapytreatments. In one embodiment, the immunotherapy treatment isbevacizumab, erlotinib, gefitinib, afatinib, or denosumab.

In one embodiment, provided herein is a method of treating, preventing,or managing renal cell carcinoma in a subject, comprising administeringto the subject a therapeutically effective amount of a compound providedherein (e.g., Compound 1), or a pharmaceutically acceptable formthereof, wherein the subject has been pre-treated or previously treatedwith one or more immunotherapy treatments. In one embodiment, theimmunotherapy treatment is bevacizumab, interleukin-2, axitinib,carfilzomib, everolimus, interferon-α, lenalidomide, pazopanib,sirolimus (rapamycin), sorafenib, sunitinib, temsirolimus, thalidomide,or tivozanib.

In one embodiment, provided herein is a method of treating, preventing,or managing bladder cancer in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., Compound 1), or a pharmaceutically acceptable form thereof,wherein the subject has been pre-treated or previously treated with oneor more immunotherapy treatments. In one embodiment, the immunotherapytreatment is Bacillus Calmette-Guérin (BCG).

In one embodiment, provided herein is a method of treating, preventing,or managing head and neck cancer in a subject, comprising administeringto the subject a therapeutically effective amount of a compound providedherein (e.g., Compound 1), or a pharmaceutically acceptable formthereof, wherein the subject has been pre-treated or previously treatedwith one or more immunotherapy treatments. In one embodiment, theimmunotherapy treatment is cetuximab, nimotuzumab, bevacizumab, orerlotinib.

In one embodiment, provided herein is a method of treating, preventing,or managing breast cancer (e.g., triple-negative breast cancer) in asubject, comprising administering to the subject a therapeuticallyeffective amount of a compound provided herein (e.g., Compound 1), or apharmaceutically acceptable form thereof, wherein the subject has beenpre-treated or previously treated with one or more immunotherapytreatments. In one embodiment, the immunotherapy treatment isbevacizumab or trastuzumab.

In one embodiment, provided herein is a method of treating, preventing,or managing ovarian cancer in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., Compound 1), or a pharmaceutically acceptable form thereof,wherein the subject has been pre-treated or previously treated with oneor more immunotherapy treatments. In one embodiment, the immunotherapytreatment is bevacizumab.

In one embodiment, provided herein is a method of treating, preventing,or managing colon cancer in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., Compound 1), or a pharmaceutically acceptable form thereof,wherein the subject has been pre-treated or previously treated with oneor more immunotherapy treatments. In one embodiment, the immunotherapytreatment is bevacizumab, cetuximab, or panitumumab.

In some embodiments, the disclosure relates to a method of treating acancer of hematopoietic origin. In certain embodiments, the cancer ofhematopoietic origin is lymphoma or leukemia. In some embodiments, thecancer of hematopoietic origin is selected from acute lymphocyticleukemia (ALL), which includes B-lineage ALL and T-lineage ALL, chroniclymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cellleukemia (HLL) and Waldenstrom's macroglobulinemia (WM); peripheral Tcell lymphomas (PTCL), adult T cell leukemia/lymphoma (ATLL), cutaneousT-cell lymphoma (CTCL), large granular lymphocytic leukemia (LGL), acutemyelocytic leukemia (AML), Hodgkin lymphoma (HL), non-Hodgkin lymphoma(NHL), follicular lymphoma, diffuse large B-cell lymphoma (DLBCL),mantle cell lymphoma (MCL), mastocytosis (e.g., systemic mastocytosis),multiple myeloma (MM), myelodysplastic syndrome (MDS),myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronicmyelogenous leukemia (CML), chronic neutrophilic leukemia (CNL), andhypereosinophilic syndrome (HES)).

In some embodiments, the disclosure relates to a method of treating asolid tumor. In some embodiments, the solid tumor is selected fromovarian cancer, colon cancer, fibrosarcoma, pancreatic cancer, lungcancer, breast cancer, lymphoma, melanoma, and glioblastoma. In someembodiment, the solid tumor is a CNS tumor. In one embodiment, the CNStumor is glioblastoma. Compound 1 shows good permeability crossblood-brain-barrier and can achieving efficacious concentration in a CNStumor.

In one embodiment, Compound 1 can be an inhibitor of P-gp(P-glycoprotein). P-glycoprotein impedes the entry of various drugs thatare used in the treatment, for example, of central nervous systemdiseases. Without being bound by a particular theory, the P-gp substratemay help maintain normal levels of P-gp activity in a patient beingtreated with a PI3K-γ inhibitor. In some embodiments, Compound 1 may notbe effluxed from a tumor and thus can maintain efficacious concentrationof Compound 1 in a tumor. For example, the concentration can bemaintained for about at least 6 hours, at least 10 hours, at least 12hours, at least 24 hours, or at least 48 hours. In one embodiment,Compound 1 can be administered once daily.

In some embodiments, Compound 1 is administered to a patient incombination with a second therapeutic that is a P-gp substrate. Inanother embodiment, Compound 1 can inhibit the efflux of the secondtherapeutic such as a cancer drug that is a P-gp substrate. Therefore,Compound 1 can be efficacious in maintaining the concentration of theco-administered cancer drug in a tumor. For example, the concentrationcan be maintained for about at least 6 hours, at least 10 hours, atleast 12 hours, at least 24 hours, or at least 48 hours. In oneembodiment, Compound 1 can be administered once daily.

P-glycoprotein is a component of the blood-brain barrier and is presenton the surface of the endothelial cells of the barrier. A PI3K-γinhibitor provided herein such as Compound 1 can be a P-glycoproteininhibitor and thus can cross the blood brain barrier. In someembodiments, a PI3K-γ inhibitor provided herein such as Compound 1 canmaintain an efficacious concentration in CNS tumor or a brain tumor(e.g., glioblastoma).

As used herein “solid tumor” refers to an abnormal mass of tissue. Solidtumors may be benign or malignant. A solid tumor grows in an anatomicalsite outside the bloodstream (in contrast, for example, to cancers ofhematopoietic origin such as leukemias) and requires the formation ofsmall blood vessels and capillaries to supply nutrients, etc. to thegrowing tumor mass. Solid tumors are named for the type of cells thatform them. Non-limiting examples of solid tumors are sarcomas,carcinomas (epithelial tumors), melanomas, and glioblastomas.

In some embodiments, the disclosure relates to a method of inhibitinggrowth of a tumor. “Inhibiting growth of a tumor” refers to slowingtumor growth and/or reducing tumor size “Inhibiting growth of a tumor”thus includes killing tumor cells as well as slowing or arresting tumorcell growth.

Exemplary solid tumors include, but are not limited to, biliary cancer(e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g.,adenocarcinoma of the breast, papillary carcinoma of the breast, mammarycancer, medullary carcinoma of the breast), brain cancer (e.g.,meningioma; glioma, e.g., astrocytoma, oligodendroglioma;medulloblastoma), cervical cancer (e.g., cervical adenocarcinoma),colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), gastric cancer (e.g., stomach adenocarcinoma),gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., headand neck squamous cell carcinoma, oral cancer (e.g., oral squamous cellcarcinoma (OSCC)), kidney cancer (e.g., nephroblastoma a.k.a. Wilms'tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular cancer(HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma,small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC),adenocarcinoma of the lung), neuroblastoma, neurofibroma (e.g.,neurofibromatosis (NF) type 1 or type 2, schwannomatosis),neuroendocrine cancer (e.g., gastroenteropancreatic neuroendoctrinetumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (e.g.,cystadenocarcinoma, ovarian embryonal carcinoma, ovarianadenocarcinoma), pancreatic cancer (e.g., pancreatic andenocarcinoma,intraductal papillary mucinous neoplasm (IPMN)), prostate cancer (e.g.,prostate adenocarcinoma), skin cancer (e.g., squamous cell carcinoma(SCC), keratoacanthoma (A), melanoma, basal cell carcinoma (BCC)) andsoft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH),liposarcoma, malignant peripheral nerve sheath tumor (MPNST),chondrosarcoma, fibrosarcoma, myxosarcoma, osteosarcoma).

In some embodiments, the disclosure relates to a method of treating acancer of hematopoietic origin comprising administering to a subject agamma selective compound (e.g., Compound 1). In certain embodiments, thecancer of hematopoietic origin is lymphoma or leukemia. In someembodiments, the cancer of hematopoietic origin is selected from acutelymphocytic leukemia (ALL), which includes B-lineage ALL and T-lineageALL, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL),hairy cell leukemia (HLL) and Waldenstrom's macroglobulinemia (WM);peripheral T cell lymphomas (PTCL), adult T cell leukemia/lymphoma(ATLL), cutaneous T-cell lymphoma (CTCL), large granular lymphocyticleukemia (LGL), acute myelocytic leukemia (AML), Hodgkin lymphoma (HL),non-Hodgkin lymphoma (NHL), follicular lymphoma, diffuse large B-celllymphoma (DLBCL), mantle cell lymphoma (MCL), mastocytosis (e.g.,systemic mastocytosis), multiple myeloma (MM), myelodysplastic syndrome(MDS), myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronicmyelogenous leukemia (CML), chronic neutrophilic leukemia (CNL), andhypereosinophilic syndrome (HES)).

In some embodiments, the disclosure relates to a method of treating asolid tumor comprising administering to a subject a gamma selectivecompound (e.g., Compound 1). In some embodiments, the solid tumor isselected from ovarian cancer, colon cancer, fibrosarcoma, pancreaticcancer, lung cancer, breast cancer, lymphoma, melanoma, andglioblastoma.

In some embodiments, the disclosure relates to a method of treating aninflammatory disease comprising administering to a subject a gammaselective compound (e.g., Compound 1).

In some embodiment, the gamma selective compound has a delta/gammaselectivity ratio of >1 to <10, 10 to <50, or 50 to <350 can be combinedwith a compound that has a gamma/delta selectivity ratio of greater thana factor of about 1, greater than a factor of about 2, greater than afactor of about 3, greater than a factor of about 5, greater than afactor of about 10, greater than a factor of about 50, greater than afactor of about 100, greater than a factor of about 200, greater than afactor of about 400, greater than a factor of about 600, greater than afactor of about 800, greater than a factor of about 1000, greater than afactor of about 1500, greater than a factor of about 2000, greater thana factor of about 5000, greater than a factor of about 10,000, orgreater than a factor of about 20,000.

Patients that can be treated with a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein, according to the methods as provided herein include, forexample, but not limited to, patients that have been diagnosed as havingpsoriasis; restenosis; atherosclerosis; ischemic stroke, BPH; breastcancer such as a ductal carcinoma, lobular carcinoma, medullarycarcinomas, colloid carcinomas, tubular carcinomas, and inflammatorybreast cancer; ovarian cancer, including epithelial ovarian tumors suchas adenocarcinoma in the ovary and an adenocarcinoma that has migratedfrom the ovary into the abdominal cavity; uterine cancer; cervicalcancer such as adenocarcinoma in the cervix epithelial includingsquamous cell carcinoma and adenocarcinomas; prostate cancer, such as aprostate cancer selected from the following: an adenocarcinoma or anadenocarcinoma that has migrated to the bone; pancreatic cancer such asepitheliod carcinoma in the pancreatic duct tissue and an adenocarcinomain a pancreatic duct; bladder cancer such as a transitional cellcarcinoma in urinary bladder, urothelial carcinomas (transitional cellcarcinomas), tumors in the urothelial cells that line the bladder,squamous cell carcinomas, adenocarcinomas, and small cell cancers;leukemia such as acute lymphoblastic leukemia, chronic myelogenousleukemia, hairy cell leukemia, myelodysplasia, myeloproliferativedisorders, NK cell leukemia (e.g., blastic plasmacytoid dendritic cellneoplasm), acute myelogenous leukemia (AML), chronic myelogenousleukemia (CML), mastocytosis, chronic lymphocytic leukemia (CLL),multiple myeloma (MM), and myelodysplastic syndrome (MDS); bone cancer;lung cancer such as non-small cell lung cancer (NSCLC), which is dividedinto squamous cell carcinomas, adenocarcinomas, and large cellundifferentiated carcinomas, and small cell lung cancer; skin cancersuch as basal cell carcinoma, melanoma, squamous cell carcinoma andactinic keratosis, which is a skin condition that sometimes developsinto squamous cell carcinoma; eye retinoblastoma; cutaneous orintraocular (eye) melanoma; primary liver cancer; kidney cancer; thyroidcancer such as papillary, follicular, medullary and anaplastic; lymphomasuch as diffuse large B-cell lymphoma, B-cell immunoblastic lymphoma, NKcell lymphoma (e.g., blastic plasmacytoid dendritic cell neoplasm), andBurkitt lymphoma; Kaposi's Sarcoma; viral-induced cancers includinghepatitis B virus (HBV), hepatitis C virus (HCV), and hepatocellularcarcinoma; human lymphotropic virus-type 1 (HTLV-1) and adult T-cellleukemia/lymphoma; and human papilloma virus (HPV) and cervical cancer;central nervous system cancers (CNS) such as primary brain tumor, whichincludes gliomas (astrocytoma, anaplastic astrocytoma, or glioblastomamultiforme), oligodendroglioma, ependymoma, meningioma, lymphoma,schwannoma, and medulloblastoma; peripheral nervous system (PNS) cancerssuch as acoustic neuromas and malignant peripheral nerve sheath tumor(MPNST) including neurofibromas and schwannomas, malignant fibrocytoma,malignant fibrous histiocytoma, malignant meningioma, malignantmesothelioma, and malignant mixed Müllerian tumor; oral cavity andoropharyngeal cancers such as, hypopharyngeal cancer, laryngeal cancer,nasopharyngeal cancer, and oropharyngeal cancer; stomach cancers such aslymphomas, gastric stromal tumors, and carcinoid tumors; testicularcancers such as germ cell tumors (GCTs), which include seminomas andnonseminomas, and gonadal stromal tumors, which include Leydig celltumors and Sertoli cell tumors; thymus cancer such as to thymomas,thymic carcinomas, Hodgkin lymphoma, non-Hodgkin lymphomas, carcinoidsor carcinoid tumors; rectal cancer; and colon cancer.

Patients that can be treated with compounds provided herein, orpharmaceutically acceptable salt, ester, prodrug, solvate, hydrate orderivative of said compounds, according to the methods provided hereininclude, for example, patients that have been diagnosed as havingconditions including, but not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma), benignmonoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma),bladder cancer, breast cancer (e.g., adenocarcinoma of the breast,papillary carcinoma of the breast, mammary cancer, medullary carcinomaof the breast), brain cancer (e.g., meningioma; glioma, e.g.,astrocytoma, oligodendroglioma; medulloblastoma), bronchus cancer,cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma,chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer,rectal cancer, colorectal adenocarcinoma), epithelial carcinoma,ependymoma, endotheliosarcoma (e.g., Kaposi's sarcoma, multipleidiopathic hemorrhagic sarcoma), endometrial cancer, esophageal cancer(e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma), Ewingsarcoma, familiar hypereosinophilia, gastric cancer (e.g., stomachadenocarcinoma), gastrointestinal stromal tumor (GIST), head and neckcancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g.,oral squamous cell carcinoma (OSCC)), heavy chain disease (e.g., alphachain disease, gamma chain disease, mu chain disease), hemangioblastoma,inflammatory myofibroblastic tumors, immunocytic amyloidosis, kidneycancer (e.g., nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma),liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma),lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer(SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung),leukemia (e.g., acute lymphoblastic leukemia (ALL), which includesB-lineage ALL and T-lineage ALL, chronic lymphocytic leukemia (CLL),prolymphocytic leukemia (PLL), hairy cell leukemia (HLL) andWaldenstrom's macroglobulinemia (WM); peripheral T cell lymphomas(PTCL), adult T cell leukemia/lymphoma (ATLL), cutaneous T-cell lymphoma(CTCL), large granular lymphocytic leukemia (LGL), acute myelocyticleukemia (AML), chronic myelocytic leukemia (CML), chronic lymphocyticleukemia (CLL)), lymphoma (e.g., Hodgkin lymphoma (HL), non-Hodgkinlymphoma (NHL), follicular lymphoma, diffuse large B-cell lymphoma(DLBCL), mantle cell lymphoma (MCL)), leiomyosarcoma (LMS), mastocytosis(e.g., systemic mastocytosis), multiple myeloma (MM), myelodysplasticsyndrome (MDS), mesothelioma, myeloproliferative disorder (MPD) (e.g.,polycythemia Vera (PV), essential thrombocytosis (ET), agnogenic myeloidmetaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathicmyelofibrosis, chronic myelogenous leukemia (CML), chronic neutrophilicleukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma,neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2,schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreaticneuroendoctrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovariancancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarianadenocarcinoma), Paget's disease of the vulva, Paget's disease of thepenis, papillary adenocarcinoma, pancreatic cancer (e.g., pancreaticandenocarcinoma, intraductal papillary mucinous neoplasm (IPMN)),pinealoma, primitive neuroectodermal tumor (PNT), prostate cancer (e.g.,prostate adenocarcinoma), rhabdomyosarcoma, retinoblastoma, salivarygland cancer, skin cancer (e.g., squamous cell carcinoma (SCC),keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small bowelcancer (e.g., appendix cancer), soft tissue sarcoma (e.g., malignantfibrous histiocytoma (MFH), liposarcoma, malignant peripheral nervesheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma),sebaceous gland carcinoma, sweat gland carcinoma, synovioma, testicularcancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer(e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma(PTC), medullary thyroid cancer), and Waldenstrom's macroglobulinemia.

Without being limited by a particular theory, in one embodiment, thecancer or disease being treated or prevented, such as a blood disorderor hematologic malignancy, has a high expression level of one or morePI3K isoform(s) (e.g., PI3K-α, PI3K-β, PI3K-δ, or PI3K-γ or acombination thereof). In one embodiment, the cancer or disease that canbe treated or prevented by methods, compositions, or kits providedherein includes a blood disorder or a hematologic malignancy, including,but not limited to, myeloid disorder, lymphoid disorder, leukemia,lymphoma, myelodysplastic syndrome (MDS), myeloproliferative disease(MPD), mast cell disorder, and myeloma (e.g., multiple myeloma), amongothers. In one embodiment, the blood disorder or the hematologicmalignancy includes, but is not limited to, acute lymphoblastic leukemia(ALL), T-cell ALL (T-ALL), B-cell ALL (B-ALL), acute myeloid leukemia(AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia(CML), blast phase CML, small lymphocytic lymphoma (SLL), CLL/SLL,transformed CLL, Richter syndrome Hodgkin lymphoma (HL), non-Hodgkinlymphoma (NHL), B-cell NHL, T-cell NHL, indolent NHL (iNHL), diffuselarge B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), aggressiveB-cell NHL, B-cell lymphoma (BCL), Richter's syndrome (RS), T-celllymphoma (TCL), peripheral T-cell lymphoma (PTCL), cutaneous T-celllymphoma (CTCL), transformed mycosis fungoides, Sézary syndrome,anaplastic large-cell lymphoma (ALCL), follicular lymphoma (FL),Waldenstrom's macroglobulinemia (WM), lymphoplasmacytic lymphoma,Burkitt lymphoma, multiple myeloma (MM), amyloidosis, MPD, essentialthrombocytosis (ET), myelofibrosis (MF), polycythemia vera (PV), chronicmyelomonocytic leukemia (CMML), myelodysplastic syndrome (MDS),angioimmunoblastic lymphoma, high-risk MDS, and low-risk MDS. In oneembodiment, the hematologic malignancy is relapsed. In one embodiment,the hematologic malignancy is refractory. In one embodiment, the canceror disease is in a pediatric patient (including an infantile patient).In one embodiment, the cancer or disease is in an adult patient.Additional embodiments of a cancer or disease being treated or preventedby methods, compositions, or kits provided herein are described hereinelsewhere.

In exemplary embodiments, the cancer or hematologic malignancy is CLL.In exemplary embodiments, the cancer or hematologic malignancy isCLL/SLL. In exemplary embodiments, the cancer or hematologic malignancyis transformed CLL or Richter syndrome. In exemplary embodiments, thecancer or hematologic malignancy is SLL. In one embodiment, withoutbeing limited by a particular theory, a compound provided herein (e.g.,a PI3K-γ selective compound provided herein) inhibits the migrationand/or activation of T-cells and myeloid cells (e.g., macrophages orpolarized M2 macrophages), reducing survival and/or proliferativesupport provided by those cells to malignant CLL cells within the tumormicroenvironment (TME). In one embodiment, without being limited by aparticular theory, the migration of CD3+T cells to the CLL-associatedchemokine CXCL12 is blocked by a compound provided herein (e.g., aPI3K-γ selective compound provided herein). In another embodiment,without being limited by a particular theory, a compound provided herein(e.g., a PI3K-γ selective compound provided herein) block the myeloidcell mediated re-growth of a cancer following chemotherapy through itseffects on inhibiting the post-chemotherapy migration of myeloid cellsinto a tumor.

In exemplary embodiments, the cancer or hematologic malignancy is iNHL.In exemplary embodiments, the cancer or hematologic malignancy is DLBCL.In exemplary embodiments, the cancer or hematologic malignancy is B-cellNHL (e.g., aggressive B-cell NHL). In exemplary embodiments, the canceror hematologic malignancy is MCL. In exemplary embodiments, the canceror hematologic malignancy is RS. In exemplary embodiments, the cancer orhematologic malignancy is AML. In exemplary embodiments, the cancer orhematologic malignancy is MM. In exemplary embodiments, the cancer orhematologic malignancy is ALL. In exemplary embodiments, the cancer orhematologic malignancy is T-ALL. In exemplary embodiments, the cancer orhematologic malignancy is B-ALL. In exemplary embodiments, the cancer orhematologic malignancy is TCL. In exemplary embodiments, the cancer orhematologic malignancy is ALCL. In exemplary embodiments, the cancer orhematologic malignancy is leukemia. In exemplary embodiments, the canceror hematologic malignancy is lymphoma. In exemplary embodiments, thecancer or hematologic malignancy is T-cell lymphoma. In exemplaryembodiments, the cancer or hematologic malignancy is MDS (e.g., lowgrade MDS). In exemplary embodiments, the cancer or hematologicmalignancy is MPD. In exemplary embodiments, the cancer or hematologicmalignancy is a mast cell disorder. In exemplary embodiments, the canceror hematologic malignancy is Hodgkin lymphoma (HL). In exemplaryembodiments, the cancer or hematologic malignancy is non-Hodgkinlymphoma. In exemplary embodiments, the cancer or hematologic malignancyis PTCL. In exemplary embodiments, the cancer or hematologic malignancyis CTCL (e.g., mycosis fungoides or Sézary syndrome). In exemplaryembodiments, the cancer or hematologic malignancy is WM. In exemplaryembodiments, the cancer or hematologic malignancy is CML. In exemplaryembodiments, the cancer or hematologic malignancy is FL. In exemplaryembodiments, the cancer or hematologic malignancy is transformed mycosisfungoides. In exemplary embodiments, the cancer or hematologicmalignancy is Sézary syndrome. In exemplary embodiments, the cancer orhematologic malignancy is acute T-cell leukemia. In exemplaryembodiments, the cancer or hematologic malignancy is acute B-cellleukemia. In exemplary embodiments, the cancer or hematologic malignancyis Burkitt lymphoma. In exemplary embodiments, the cancer or hematologicmalignancy is myeloproliferative neoplasms. In exemplary embodiments,the cancer or hematologic malignancy is splenic marginal zone. Inexemplary embodiments, the cancer or hematologic malignancy is nodalmarginal zone. In exemplary embodiments, the cancer or hematologicmalignancy is extranodal marginal zone.

In one embodiment, the cancer or hematologic malignancy is a B celllymphoma. In a specific embodiment, provided herein is a method oftreating or managing a B cell lymphoma comprising administering to apatient a therapeutically effective amount of a compound providedherein, or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof. Also provided herein is a method of treating orlessening one or more of the symptoms associated with a B cell lymphomacomprising administering to a patient a therapeutically effective amountof a compound provided herein, or a pharmaceutically acceptablederivative (e.g., salt or solvate) thereof. In one embodiment, the Bcell lymphoma is iNHL. In another embodiment, the B cell lymphoma isfollicular lymphoma. In another embodiment, the B cell lymphoma isWaldenstrom's macroglobulinemia (lymphoplasmacytic lymphoma). In anotherembodiment, the B cell lymphoma is marginal zone lymphoma (MZL). Inanother embodiment, the B cell lymphoma is MCL. In another embodiment,the B cell lymphoma is HL. In another embodiment, the B cell lymphoma isiNHL. In another embodiment, the B cell lymphoma is DLBCL. In anotherembodiment, the B cell lymphoma is Richter's lymphoma.

In one embodiment, the cancer or hematologic malignancy is a T celllymphoma. In a specific embodiment, provided herein is a method oftreating or managing a T cell lymphoma comprising administering to apatient a therapeutically effective amount of a compound providedherein, or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof. Also provided herein is a method of treating orlessening one or more of the symptoms associated with a T cell lymphomacomprising administering to a patient a therapeutically effective amountof a compound provided herein, or a pharmaceutically acceptablederivative (e.g., salt or solvate) thereof. In one embodiment, the Tcell lymphoma is peripheral T cell lymphoma (PTCL). In anotherembodiment, the T cell lymphoma is cutaneous T cell lymphoma (CTCL).

In one embodiment, the cancer or hematologic malignancy is Sézarysyndrome. In a specific embodiment, provided herein is a method oftreating or managing Sézary syndrome comprising administering to apatient a therapeutically effective amount of a compound providedherein, or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof. Also provided herein is a method of treating orlessening one or more of the symptoms associated with Sézary syndromecomprising administering to a patient a therapeutically effective amountof a compound provided herein, or a pharmaceutically acceptablederivative (e.g., salt or solvate) thereof. The symptoms associated withSézary syndrome include, but are not limited to, epidermotropism byneoplastic CD4+ lymphocytes, Pautrier's microabscesses, erythroderma,lymphadenopathy, atypical T cells in the peripheral blood, andhepatosplenomegalyIn one embodiment, the therapeutically effectiveamount for treating or managing Sézary syndrome is from about 25 mg to75 mg, administered twice daily. In other embodiments, thetherapeutically effective amount is from about 50 mg to about 75 mg,from about 30 mg to about 65 mg, from about 45 mg to about 60 mg, fromabout 30 mg to about 50 mg, or from about 55 mg to about 65 mg, each ofwhich is administered twice daily. In one embodiment, the effectiveamount is about 60 mg, administered twice daily.

In one embodiment, the cancer or hematologic malignancy is relapsed. Inone embodiment, the cancer or hematologic malignancy is refractory. Incertain embodiments, the cancer being treated or prevented is a specificsub-type of cancer described herein. In certain embodiments, thehematologic malignancy being treated or prevented is a specific sub-typeof hematologic malignancy described herein. Certain classifications oftype or sub-type of a cancer or hematologic malignancy provided hereinis known in the art. Without being limited by a particular theory, it isbelieved that many of the cancers that become relapsed or refractorydevelop resistance to the particular prior therapy administered to treatthe cancers. Thus, without being limited by a particular theory, acompound provided herein can provide a second line therapy by providingan alternative mechanism to treat cancers different from thosemechanisms utilized by certain prior therapies. Accordingly, in oneembodiment, provided herein is a method of treating or managing canceror hematologic malignancy comprising administering to a patient atherapeutically effective amount of a compound provided herein, or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,wherein the cancer or hematologic malignancy is relapsed after, orrefractory to, a prior therapy.

In exemplary embodiments, the cancer or hematologic malignancy isrefractory iNHL. In exemplary embodiments, the cancer or hematologicmalignancy is refractory CLL. In exemplary embodiments, the cancer orhematologic malignancy is refractory SLL. In exemplary embodiments, thecancer or hematologic malignancy is refractory to rituximab therapy. Inexemplary embodiments, the cancer or hematologic malignancy isrefractory to chemotherapy. In exemplary embodiments, the cancer orhematologic malignancy is refractory to radioimmunotherapy (RIT). Inexemplary embodiments, the cancer or hematologic malignancy is iNHL, FL,splenic marginal zone, nodal marginal zone, extranodal marginal zone, orSLL, the cancer or hematologic malignancy is refractory to rituximabtherapy, chemotherapy, and/or RIT.

In another exemplary embodiment, the cancer or hematologic malignancy islymphoma, and the cancer is relapsed after, or refractory to, thetreatment by a BTK inhibitor such as, but not limited to, ibrutinib orONO-4059. In another exemplary embodiment, the cancer or hematologicmalignancy is CLL, and the cancer is relapsed after, or refractory to,the treatment by a BTK inhibitor such as, but not limited to, ibrutiniband AVL-292.

In certain embodiments, provided herein are methods of treating orpreventing a solid tumor in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., a selective PI3K-γ inhibitor, e.g., Compound 1), or apharmaceutically acceptable form thereof.

In one embodiment, the solid tumor is selected from one or more of: acancer of the pulmonary system, a brain cancer, a cancer of thegastrointestinal tract, a skin cancer, a genitourinary cancer, apancreatic cancer, a lung cancer, a medulloblastoma, a basal cellcarcinoma, a glioma, a breast cancer, a prostate cancer, a testicularcancer, an esophageal cancer, a hepatocellular cancer, a gastric cancer,a gastrointestinal stromal tumor (GIST), a colon cancer, a colorectalcancer, an ovarian cancer, a melanoma, a neuroectodermal tumor, head andneck cancer, a sarcoma, a soft-tissue sarcoma, fibrosarcoma,myxosarcoma, liposarcoma, a chondrosarcoma, an osteogenic sarcoma, achordoma, an angiosarcoma, an endotheliosarcoma, a lymphangiosarcoma, alymphangioendotheliosarcoma, a synovioma, a mesothelioma, aleiomyosarcoma, a cervical cancer, a uterine cancer, an endometrialcancer, a carcinoma, a bladder carcinoma, an epithelial carcinoma, asquamous cell carcinoma, an adenocarcinoma, a bronchogenic carcinoma, arenal cell carcinoma, a hepatoma, a bile duct carcinoma, aneuroendocrine cancer, a carcinoid tumor, diffuse type giant cell tumor,and glioblastoma.

In one embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered at a dose sufficient to cause a decrease intumor growth of at least 10%, 20%, 30%, 40%, or 50% compared to areference value, after administration of the compound.

In one embodiment, the method further comprises administering animmunomodulator to the subject. In one embodiment, the immunomodulatoris a PDL-1 inhibitor or an anti-PDL-1 antibody.

In one embodiment, the method further comprises administering aPI3K-delta inhibitor to the subject.

In one embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered at a dose such that the level of the compoundin the subject is higher than the compound's IC50 of PI3K-gammainhibition during at least 70%, 80%, 90%, 95%, 97%, 98%, or 99% of aselected time period, e.g., 6 hours, 12 hours, 24 hours, or 48 hours,immediately following the administration. In one embodiment, thecompound, or a pharmaceutically acceptable form thereof, is administeredat a dose such that the level of the compound in the subject is lowerthan the compound's IC50 of PI3K-delta inhibition during at least 70%,80%, 90%, 95%, 97%, 98%, or 99% of a selected time period, e.g., 6hours, 12 hours, 24 hours, or 48 hours, immediately following theadministration. In one embodiment, the level of the compound is measuredfrom the subject's plasma. In one embodiment, the level of the compoundis measured from the subject's tissue.

In one embodiment, the subject has been previously treated withcyclophosphamide, docetaxel, paclitaxel, 5-FU, or temozolomide.

In one embodiment, the anti-tumor effect of the compound is maintainedfor a period of time after the discontinuation of treatment with thecompound. In one embodiment, the period of time is at least 1 day, 2days, 3 days, 4 days, 5 days, or 6 days.

Treatment of an Inflammatory Disorder

In one embodiment, provided herein is a method of treating aninflammation disorder, including autoimmune diseases in a subject. Themethod comprises administering to said subject a therapeuticallyeffective amount of a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein. Examples ofautoimmune diseases include but are not limited to acute disseminatedencephalomyelitis (ADEM), Addison's disease, antiphospholipid antibodysyndrome (APS), aplastic anemia, autoimmune hepatitis, autoimmune skindisease, coeliac disease, Crohn's disease, Diabetes mellitus (type 1),Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome (GBS),Hashimoto's disease, lupus erythematosus, multiple sclerosis, myastheniagravis, opsoclonus myoclonus syndrome (OMS), optic neuritis, Ord'sthyroiditis, oemphigus, polyarthritis, primary biliary cirrhosis,psoriasis, rheumatoid arthritis, Reiter's syndrome, Takayasu'sarteritis, temporal arteritis (also known as “giant cell arteritis”),warm autoimmune hemolytic anemia, Wegener's granulomatosis, alopeciauniversalis (e.g., inflammatory alopecia), Chagas disease, chronicfatigue syndrome, dysautonomia, endometriosis, hidradenitis suppurativa,interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma,ulcerative colitis, vitiligo, and vulvodynia. Other disorders includebone-resorption disorders and thrombosis.

Inflammation takes on many forms and includes, but is not limited to,acute, adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse,disseminated, exudative, fibrinous, fibrosing, focal, granulomatous,hyperplastic, hypertrophic, interstitial, metastatic, necrotic,obliterative, parenchymatous, plastic, productive, proliferous,pseudomembranous, purulent, sclerosing, seroplastic, serous, simple,specific, subacute, suppurative, toxic, traumatic, and/or ulcerativeinflammation.

Exemplary inflammatory conditions include, but are not limited to,inflammation associated with acne, anemia (e.g., aplastic anemia,haemolytic autoimmune anaemia), asthma, arteritis (e.g., polyarteritis,temporal arteritis, periarteritis nodosa, Takayasu's arteritis),arthritis (e.g., crystalline arthritis, osteoarthritis, psoriaticarthritis, gout flare, gouty arthritis, reactive arthritis, rheumatoidarthritis and Reiter's arthritis), ankylosing spondylitis, amylosis,amyotrophic lateral sclerosis, autoimmune diseases, allergies orallergic reactions, atherosclerosis, bronchitis, bursitis, chronicprostatitis, conjunctivitis, Chagas disease, chronic obstructivepulmonary disease, cermatomyositis, diverticulitis, diabetes (e.g., typeI diabetes mellitus, type 2 diabetes mellitus), a skin condition (e.g.,psoriasis, eczema, burns, dermatitis, pruritus (itch)), endometriosis,Guillain-Barre syndrome, infection, ischaemic heart disease, Kawasakidisease, glomerulonephritis, gingivitis, hypersensitivity, headaches(e.g., migraine headaches, tension headaches), ileus (e.g.,postoperative ileus and ileus during sepsis), idiopathicthrombocytopenic purpura, interstitial cystitis (painful bladdersyndrome), gastrointestinal disorder (e.g., selected from peptic ulcers,regional enteritis, diverticulitis, gastrointestinal bleeding,eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis,eosinophilic gastritis, eosinophilic gastroenteritis, eosinophiliccolitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD, orits synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn'sdisease, ulcerative colitis, collagenous colitis, lymphocytic colitis,ischaemic colitis, diversion colitis, Behcet's syndrome, indeterminatecolitis) and inflammatory bowel syndrome (IBS)), lupus, multiplesclerosis, morphea, myeasthenia gravis, myocardial ischemia, nephroticsyndrome, pemphigus vulgaris, pernicious aneaemia, peptic ulcers,polymyositis, primary biliary cirrhosis, neuroinflammation associatedwith brain disorders (e.g., Parkinson's disease, Huntington's disease,and Alzheimer's disease), prostatitis, chronic inflammation associatedwith cranial radiation injury, pelvic inflammatory disease, polymyalgiarheumatic, reperfusion injury, regional enteritis, rheumatic fever,systemic lupus erythematosus, scleroderma, scierodoma, sarcoidosis,spondyloarthopathies, Sjogren's syndrome, thyroiditis, transplantationrejection, tendonitis, trauma or injury (e.g., frostbite, chemicalirritants, toxins, scarring, burns, physical injury), vasculitis,vitiligo and Wegener's granulomatosis. In certain embodiments, theinflammatory disorder is selected from arthritis (e.g., rheumatoidarthritis), inflammatory bowel disease, inflammatory bowel syndrome,asthma, psoriasis, endometriosis, interstitial cystitis andprostatistis. In certain embodiments, the inflammatory condition is anacute inflammatory condition (e.g., for example, inflammation resultingfrom infection). In certain embodiments, the inflammatory condition is achronic inflammatory condition (e.g., conditions resulting from asthma,arthritis and inflammatory bowel disease). The compounds can also beuseful in treating inflammation associated with trauma andnon-inflammatory myalgia.

Immune disorders, such as auto-immune disorders, include, but are notlimited to, arthritis (including rheumatoid arthritis,spondyloarthopathies, gouty arthritis, degenerative joint diseases suchas osteoarthritis, systemic lupus erythematosus, Sjogren's syndrome,ankylosing spondylitis, undifferentiated spondylitis, Behcet's disease,haemolytic autoimmune anaemias, multiple sclerosis, amyotrophic lateralsclerosis, amylosis, acute painful shoulder, psoriatic, and juvenilearthritis), asthma, atherosclerosis, osteoporosis, bronchitis,tendonitis, bursitis, skin condition (e.g., psoriasis, eczema, burns,dermatitis, pruritus (itch)), enuresis, eosinophilic disease,gastrointestinal disorder (e.g., selected from peptic ulcers, regionalenteritis, diverticulitis, gastrointestinal bleeding, eosinophilicgastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilicgastritis, eosinophilic gastroenteritis, eosinophilic colitis),gastritis, diarrhea, gastroesophageal reflux disease (GORD, or itssynonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn's disease,ulcerative colitis, collagenous colitis, lymphocytic colitis, ischaemiccolitis, diversion colitis, Behcet's syndrome, indeterminate colitis)and inflammatory bowel syndrome (IBS)), relapsing polychondritis (e.g.,atrophic polychondritis and systemic polychondromalacia), and disordersameliorated by a gastroprokinetic agent (e.g., ileus, postoperativeileus and ileus during sepsis; gastroesophageal reflux disease (GORD, orits synonym GERD); eosinophilic esophagitis, gastroparesis such asdiabetic gastroparesis; food intolerances and food allergies and otherfunctional bowel disorders, such as non-ulcerative dyspepsia (NUD) andnon-cardiac chest pain (NCCP, including costo-chondritis)). In certainembodiments, a method of treating inflammatory or autoimmune diseases isprovided comprising administering to a subject (e.g., a mammal) atherapeutically effective amount of a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein, that selectively inhibit PI3K-δ and/or PI3K-γ as compared to allother type I PI3 kinases. Such selective inhibition of PI3K-δ and/orPI3K-γ can be advantageous for treating any of the diseases orconditions described herein. For example, selective inhibition of PI3K-δand/or PI3K-γ can inhibit inflammatory responses associated withinflammatory diseases, autoimmune disease, or diseases related to anundesirable immune response including, but not limited to asthma,emphysema, allergy, dermatitis, rheumatoid arthritis, psoriasis, lupuserythematosus, anaphylaxsis, or graft versus host disease. Selectiveinhibition of PI3K-δ and/or PI3K-γ can further provide for a reductionin the inflammatory or undesirable immune response without a concomitantreduction in the ability to reduce a bacterial, viral, and/or fungalinfection. Selective inhibition of both PI3K-δ and PI3K-γ can beadvantageous for inhibiting the inflammatory response in the subject toa greater degree than that would be provided for by inhibitors thatselectively inhibit PI3K-6 or PI3K-γ alone. In one aspect, one or moreof the subject methods are effective in reducing antigen specificantibody production in vivo by about 2-fold, 3-fold, 4-fold, 5-fold,7.5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 250-fold, 500-fold,750-fold, or about 1000-fold or more. In another aspect, one or more ofthe subject methods are effective in reducing antigen specific IgG3and/or IgGM production in vivo by about 2-fold, 3-fold, 4-fold, 5-fold,7.5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 250-fold, 500-fold,750-fold, or about 1000-fold or more.

In one aspect, one or more of the subject methods are effective inameliorating symptoms associated with rheumatoid arthritis including,but not limited to a reduction in the swelling of joints, a reduction inserum anti-collagen levels, and/or a reduction in joint pathology suchas bone resorption, cartilage damage, pannus, and/or inflammation. Inanother aspect, the subject methods are effective in reducing ankleinflammation by at least about 2%, 5%, 10%, 15%, 20%, 25%, 30%, 50%, or60%, or about 75% to 90%. In another aspect, the subject methods areeffective in reducing knee inflammation by at least about 2%, 5%, 10%,15%, 20%, 25%, 30%, 50%, or 60%, or about 75% to 90% or more. In stillanother aspect, the subject methods are effective in reducing serumanti-type II collagen levels by at least about 10%, 12%, 15%, 20%, 24%,25%, 30%, 35%, 50%, 60%, 75%, 80%, 86%, or 87%, or about 90% or more. Inanother aspect, the subject methods are effective in reducing anklehistopathology scores by about 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%,60%, 75%, 80%, or 90%, or more. In still another aspect, the subjectmethods are effective in reducing knee histopathology scores by about5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 75%, 80%, or 90%, or more.

In certain embodiments, provided herein are methods of treating orpreventing arthritis in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., a selective PI3K-γ inhibitor, e.g., Compound 1), or apharmaceutically acceptable form thereof. In one embodiment, thetreatment results in reduction of periosteal bone formation in thesubject. In one embodiment, the treatment results in at least 10%, 20%,40%, 47%, 50%, 52%, 60%, 80%, or 82% reduction of periosteal boneformation in the subject, compared to a reference value. In oneembodiment, the periosteal bone formation is measured by histopathologyscore or periosteal bone width. In one embodiment, the treatment resultsin at least 10%, 20%, 27%, 30%, 36%, 40%, 45%, 50%, or 57% reduction ofinflammation, at least 10%, 20%, 28%, 30%, 40%, 44%, 50%, or 60%, 70%,or 71% reduction of pannus, at least 10%, 20%, 28%, 30%, 40%, 45%, 50%,or 59% reduction of cartilage damage, or at least 10%, 20%, 25%, 30%,40%, 44%, 50%, 60%, or 65% reduction of bone resorption in the subject,compared to a reference value. In one embodiment, wherein the treatmentresults in reduction of joint swelling or anti-collagen level in thesubject.

In some embodiments, provided herein are methods for treating disordersor conditions in which the 6 isoform of PI3K is implicated to a greaterextent than other PI3K isoforms such as PI3K-α and/or PI3K-β. In someembodiments, provided herein are methods for treating disorders orconditions in which the γ isoform of PI3K is implicated to a greaterextent than other PI3K isoforms such as PI3K-α and/or PI3K-β. Selectiveinhibition of PI3K-δ and/or PI3K-γ can provide advantages over usingless selective compounds which inhibit PI3K-α and/or PI3K-β, such as animproved side effects profile or lessened reduction in the ability toreduce a bacterial, viral, and/or fungal infection.

In other embodiments, provided herein are methods of using a compoundprovided herein, or a pharmaceutically acceptable form (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein, to treat respiratorydiseases including, but not limited to, diseases affecting the lobes oflung, pleural cavity, bronchial tubes, trachea, upper respiratory tract,or the nerves and muscle for breathing. For example, methods areprovided to treat obstructive pulmonary disease. Chronic obstructivepulmonary disease (COPD) is an umbrella term for a group of respiratorytract diseases that are characterized by airflow obstruction orlimitation. Conditions included in this umbrella term include, but arenot limited to: chronic bronchitis, emphysema, and bronchiectasis.

In another embodiment, a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein is used forthe treatment of asthma. Also, a compound provided herein, or apharmaceutically acceptable form thereof, or a pharmaceuticalcomposition described herein, can be used for the treatment ofendotoxemia and sepsis. In one embodiment, the compounds orpharmaceutical compositions described herein are used to for thetreatment of rheumatoid arthritis (RA). In yet another embodiment, thecompounds or pharmaceutical compositions described herein is used forthe treatment of contact or atopic dermatitis. Contact dermatitisincludes irritant dermatitis, phototoxic dermatitis, allergicdermatitis, photoallergic dermatitis, contact urticaria, systemiccontact-type dermatitis and the like. Irritant dermatitis can occur whentoo much of a substance is used on the skin of when the skin issensitive to certain substance. Atopic dermatitis, sometimes calledeczema, is a kind of dermatitis, an atopic skin disease.

In certain embodiments, provided herein are methods of reducingneutrophil migration or infiltration in a subject suffering from aninflammatory disease, comprising administering to the subject atherapeutically effective amount of a compound provided herein (e.g., aselective PI3K-γ inhibitor, e.g., Compound 1), or a pharmaceuticallyacceptable form thereof. In one embodiment, the neutrophil migration orinfiltration is reduced by at least about 10%, 20%, 40%, 60%, 80%, or90% compared to a reference value, after administration of the compound.In one embodiment, the inflammatory disease is selected from the groupconsisting of COPD, arthritis, asthma, psoriasis, scleroderma, myositis,sarcoidosis, dermatomyositis, CREST syndrome, systemic lupuserythematosus, Sjorgren syndrome, encephalomyelitis, and inflammatorybowel disease (IBD). In one embodiment, the inflammatory disease is COPDor arthritis. In one embodiment, the subject is unresponsive orrefractory to a PI3K-delta inhibitor treatment.

Treatment of Other Disorders or Conditions

In some embodiments, the disclosure provides a method of treatingdiseases related to vasculogenesis or angiogenesis in a subject thatcomprises administering to said subject a therapeutically effectiveamount of a compound provided herein, or a pharmaceutically acceptableform (e.g., pharmaceutically acceptable salts, hydrates, solvates,isomers, prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein. In some embodiments, saidmethod is for treating a disease selected from tumor angiogenesis,chronic inflammatory disease such as rheumatoid arthritis and chronicinflammatory demyelinating polyneuropathy, atherosclerosis, inflammatorybowel disease, skin diseases such as psoriasis, eczema, and scleroderma,diabetes, diabetic retinopathy, retinopathy of prematurity, age-relatedmacular degeneration, hemangioma, glioma, melanoma, Kaposi's sarcoma andovarian, breast, lung, pancreatic, prostate, colon and epidermoidcancer.

In addition, the compounds described herein can be used for thetreatment of arteriosclerosis, including atherosclerosis.Arteriosclerosis is a general term describing any hardening of medium orlarge arteries. Atherosclerosis is a hardening of an artery specificallydue to an atheromatous plaque.

In some embodiments, provided herein is a method of treating acardiovascular disease in a subject that comprises administering to saidsubject a therapeutically effective amount of a compound providedherein, or a pharmaceutically acceptable form (e.g., pharmaceuticallyacceptable salts, hydrates, solvates, isomers, prodrugs, andisotopically labeled derivatives) thereof, or a pharmaceuticalcomposition as provided herein. Examples of cardiovascular conditionsinclude, but are not limited to, atherosclerosis, restenosis, vascularocclusion and carotid obstructive disease.

In some embodiments, the disclosure relates to a method of treatingdiabetes in a subject that comprises administering to said subject atherapeutically effective amount of a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein.

In addition, a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein, can be usedto treat acne. In certain embodiments, the inflammatory condition and/orimmune disorder is a skin condition. In some embodiments, the skincondition is pruritus (itch), psoriasis, eczema, burns or dermatitis. Incertain embodiments, the skin condition is psoriasis. In certainembodiments, the skin condition is pruritis.

In certain embodiments, the inflammatory disorder and/or the immunedisorder is a gastrointestinal disorder. In some embodiments, thegastrointestinal disorder is selected from gastrointestinal disorder(e.g., selected from peptic ulcers, regional enteritis, diverticulitis,gastrointestinal bleeding, eosinophilic gastrointestinal disorders(e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilicgastroenteritis, eosinophilic colitis), gastritis, diarrhea,gastroesophageal reflux disease (GORD, or its synonym GERD),inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerativecolitis, collagenous colitis, lymphocytic colitis, ischaemic colitis,diversion colitis, Behcet's syndrome, indeterminate colitis) andinflammatory bowel syndrome (IBS)). In certain embodiments, thegastrointestinal disorder is inflammatory bowel disease (IBD).

Further, a compound provided herein, or a pharmaceutically acceptableform (e.g., pharmaceutically acceptable salts, hydrates, solvates,isomers, prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein, can be used for thetreatment of glomerulonephritis. Glomerulonephritis is a primary orsecondary autoimmune renal disease characterized by inflammation of theglomeruli. It can be asymptomatic, or present with hematuria and/orproteinuria. There are many recognized types, divided in acute, subacuteor chronic glomerulonephritis. Causes are infectious (bacterial, viralor parasitic pathogens), autoimmune or paraneoplastic.

In some embodiments, provided herein are compounds, or pharmaceuticallyacceptable forms (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or pharmaceutical compositions as provided herein, for thetreatment of multiorgan failure. Also provided herein are compounds, orpharmaceutically acceptable forms (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,for the treatment of liver diseases (including diabetes), gall bladderdisease (including gallstones), pancreatitis or kidney disease(including proliferative glomerulonephritis and diabetes-induced renaldisease) or pain in a subject.

In some embodiments, provided herein are compounds, or pharmaceuticallyacceptable forms (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or pharmaceutical compositions as provided herein, for theprevention of blastocyte implantation in a subject.

In some embodiments, provided herein are compounds, or pharmaceuticallyacceptable forms (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or pharmaceutical compositions as provided herein, for thetreatment of disorders involving platelet aggregation or plateletadhesion, including, but not limited to, Idiopathic thrombocytopenicpurpura, Bernard-Soulier syndrome, Glanzmann's thrombasthenia, Scott'ssyndrome, von Willebrand disease, Hermansky-Pudlak Syndrome, and Grayplatelet syndrome.

In some embodiments, provided herein are compounds, or pharmaceuticallyacceptable forms (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or pharmaceutical compositions as provided herein, for thetreatment of a disease which is skeletal muscle atrophy, skeletal ormuscle hypertrophy. In some embodiments, provided herein are compounds,or pharmaceutically acceptable forms (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,for the treatment of disorders that include, but are not limited to,cancers as discussed herein, transplantation-related disorders (e.g.,lowering rejection rates, graft-versus-host disease, etc.), muscularsclerosis (MS), allergic disorders (e.g., arthritis, allergicencephalomyelitis) and other immunosuppressive-related disorders,metabolic disorders (e.g., diabetes), reducing intimal thickeningfollowing vascular injury, and misfolded protein disorders (e.g.,Alzheimer's Disease, Gaucher's Disease, Parkinson's Disease,Huntington's Disease, cystic fibrosis, macular degeneration, retinitispigmentosa, and prion disorders) (as mTOR inhibition can alleviate theeffects of misfolded protein aggregates). The disorders also includehamartoma syndromes, such as tuberous sclerosis and Cowden Disease (alsotermed Cowden syndrome and multiple hamartoma syndrome).

Additionally, a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein, can be usedfor the treatment of bursitis, lupus, acute disseminatedencephalomyelitis (ADEM), Addison's disease, antiphospholipid antibodysyndrome (APS), amyloidosis (including systemic and localizedamyloidosis; and primary and secondary amyloidosis), aplastic anemia,autoimmune hepatitis, coeliac disease, crohn's disease, diabetesmellitus (type 1), eosinophilic gastroenterides, goodpasture's syndrome,graves' disease, guillain-barré syndrome (GBS), hashimoto's disease,inflammatory bowel disease, lupus erythematosus (including cutaneouslupus erythematosus and systemic lupus erythematosus), myastheniagravis, opsoclonus myoclonus syndrome (OMS), optic neuritis, ord'sthyroiditis, ostheoarthritis, uveoretinitis, pemphigus, polyarthritis,primary biliary cirrhosis, reiter's syndrome, takayasu's arteritis,temporal arteritis, warm autoimmune hemolytic anemia, wegener'sgranulomatosis, alopecia universalis, chagas' disease, chronic fatiguesyndrome, dysautonomia, endometriosis, hidradenitis suppurativa,interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma,ulcerative colitis, vitiligo, vulvodynia, appendicitis, arteritis,arthritis, blepharitis, bronchiolitis, bronchitis, cervicitis,cholangitis, cholecystitis, chorioamnionitis, colitis, conjunctivitis,cystitis, dacryoadenitis, dermatomyositis, endocarditis, endometritis,enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis,fibrositis, gastritis, gastroenteritis, gingivitis, hepatitis,hidradenitis, ileitis, iritis, laryngitis, mastitis, meningitis,myelitis, myocarditis, myositis, nephritis, omphalitis, oophoritis,orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis,peritonitis, pharyngitis, pleuritis, phlebitis, pneumonitis, proctitis,prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis,stomatitis, synovitis, pigmented villonodular synovitis (also known astenosynovial giant cell tumor), tendonitis, tonsillitis, uveitis (e.g.,ocular uveitis), vaginitis, vasculitis, or vulvitis.

Further, the compounds provided herein may be used for the treatment ofPerennial allergic rhinitis, Mesenteritis, Peritonitis, Acrodermatitis,Angiodermatitis, Atopic dermatitis, Contact dermatitis, Eczema, Erythemamultiforme, Intertrigo, Stevens Johnson syndrome, Toxic epidermalnecrolysis, Skin allergy, Severe allergic reaction/anaphylaxis, Allergicgranulomatosis, Wegener granulomatosis, Allergic conjunctivitis,Chorioretinitis, Conjunctivitis, Infectious keratoconjunctivitis,Keratoconjunctivitis, Ophthalmia neonatorum, Trachoma, Uveitis, Ocularinflammation, ocular lymphoma, MALT lymphoma, Blepharoconjunctivitis,Mastitis, Gingivitis, Pericoronitis, Pharyngitis, Rhinopharyngitis,Sialadenitis, Musculoskeletal system inflammation, Adult onset Stillsdisease, Behcets disease, Bursitis, Chondrocalcinosis, Dactylitis, Feltysyndrome, Gout, Infectious arthritis, Lyme disease, Inflammatoryosteoarthritis, Periarthritis, Reiter syndrome, Ross River virusinfection, Acute Respiratory, Distress Syndrome, Acute bronchitis, Acutesinusitis, Allergic rhinitis, Asthma, Severe refractory asthma,Pharyngitis, Pleurisy, Rhinopharyngitis, Seasonal allergic rhinitis,Sinusitis, Status asthmaticus, Tracheobronchitis, Rhinitis, Serositis,Meningitis, Neuromyelitis optica, Poliovirus infection, Alport syndrome,Balanitis, Epididymitis, Epididymo orchitis, Focal segmental,Glomerulosclerosis, Glomerulonephritis, IgA Nephropathy (Berger'sDisease), Orchitis, Parametritis, Pelvic inflammatory disease,Prostatitis, Pyelitis, Pyelocystitis, Pyelonephritis, Wegenergranulomatosis, Hyperuricemia, Aortitis, Arteritis, Chylopericarditis,Dressler syndrome, Endarteritis, Endocarditis, Extracranial temporalarteritis, HIV associated arteritis, Intracranial temporal arteritis,Kawasaki disease, Lymphangiophlebitis, Mondor disease, Periarteritis, orPericarditis.

In other aspects, the compounds provided herein are used for thetreatment of Autoimmune hepatitis, Jejunitis, Mesenteritis, Mucositis,Nonalcoholic steatohepatitis, Non-viral hepatitis, Autoimmunepancreatitis, Perihepatitis, Peritonitis, Pouchitis, Proctitis,Pseudomembranous colitis, Rectosigmoiditis, Salpingoperitonitis,Sigmoiditis, Steatohepatitis, Ulcerative colitis, Churg Strausssyndrome, Ulcerative proctitis, Irritable bowel syndrome,Gastrointestinal inflammation, Acute enterocolitis, Anusitis, Balsernecrosis, Cholecystitis, Colitis, Crohn's disease, Diverticulitis,Enteritis, Enterocolitis, Enterohepatitis, Eosinophilic esophagitis,Esophagitis, Gastritis, Hemorrhagic enteritis, Hepatitis, Hepatitisvirus infection, Hepatocholangitis, Hypertrophic gastritis, Ileitis,Ileocecitis, Sarcoidosis, Inflammatory bowel disease, Ankylosingspondylitis, Rheumatoid arthritis, Juvenile rheumatoid arthritis,Psoriasis, Psoriatic arthritis, Lupus (cutaneous/systemic/nephritis),AIDS, Agammaglobulinemia, AIDS related complex, Bruton's disease,Chediak Higashi syndrome, Common variable immunodeficiency, DiGeorgesyndrome, Dysgammaglobulinemia, Immunoglobulindeficiency, Job syndrome,Nezelof syndrome, Phagocyte bactericidal disorder, Wiskott Aldrichsyndrome, Asplenia, Elephantiasis, Hypersplenism, Kawasaki disease,Lymphadenopathy, Lymphedema, Lymphocele, Nonne Milroy Meige syndrome,Spleen disease, Splenomegaly, Thymoma, Thymus disease, Perivasculitis,Phlebitis, Pleuropericarditis, Polyarteritis nodosa, Vasculitis,Takayasus arteritis, Temporal arteritis, Thromboangiitis,Thromboangiitis obliterans, Thromboendocarditis, Thrombophlebitis, orCOPD.

In another aspect, provided herein are methods of disrupting thefunction of a leukocyte or disrupting a function of an osteoclast. Themethod includes contacting the leukocyte or the osteoclast with afunction disrupting amount of a compound provided herein.

In another aspect, provided herein are methods for the treatment of anophthalmic disease by administering one or more of compounds providedherein, or pharmaceutically acceptable forms thereof, or pharmaceuticalcompositions as provided herein, to the eye of a subject.

Methods are further provided for administering the compounds providedherein via eye drop, intraocular injection, intravitreal injection,topically, or through the use of a drug eluting device, microcapsule,implant, or microfluidic device. In some cases, the compounds providedherein are administered with a carrier or excipient that increases theintraocular penetrance of the compound such as an oil and water emulsionwith colloid particles having an oily core surrounded by an interfacialfilm.

In certain embodiments, provided herein are methods of treating,preventing, and/or managing a disease or a disorder using a compound, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,wherein the disease or disorder is: Crohn's disease; cutaneous lupus;multiple sclerosis; rheumatoid arthritis; and systemic lupuserythematosus.

In other embodiments, provided herein are methods of treating,preventing and/or managing a disease or a disorder using a compound, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,wherein the disease or disorder is: ankylosing spondylitis; chronicobstructive pulmonary disease; myasthenia gravis; ocular uveitis,psoriasis; and psoriatic arthritis.

In other embodiments, provided herein are methods of treating,preventing and/or managing a disease or a disorder using a compound, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,wherein the disease or disorder is: adult-onset Still's disease;inflammatory alopecia; amyloidosis; antiphospholipid syndrome;autoimmune hepatitis; autoimmune skin disease, Behcet's disease; chronicinflammatory demyelinating polyneuropathy; eosinophilic gastroenteritis;inflammatory myopathies, pemphigus, polymyalgia rheumatica; relapsingpolychondritis; Sjogren's syndrome; temporal arthritis; ulcerativecolitis; vasculis; vitiligo, and Wegner's granulomatosis.

In other embodiments, provided herein are methods of treating,preventing and/or managing a disease or a disorder using a compound, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,wherein the disease or disorder is: gout flare; sarcoidosis; andsystemic sclerosis.

In certain embodiments, provided herein are methods of treating,preventing and/or managing a disease or a disorder using a compound, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or pharmaceutical compositions as provided herein,wherein the disease or disorder is: asthma; arthritis (e.g., rheumatoidarthritis and psoriatic arthritis); psoriasis; scleroderma; myositis(e.g., dermatomyositis); lupus (e.g., cutaneous lupus erythematosus(“CLE”) or systemic lupus erythematosus (“SLE”)); or Sjögren's syndrome.

Efficacy of a compound provided herein in treating, preventing and/ormanaging the disease or disorder can be tested using various animalmodels known in the art. For example: efficacy in treating, preventingand/or managing asthma can be assessed using ova induced asthma modeldescribed, for example, in Lee et al. (2006) J Allergy Clin Immunol118(2):403-9; efficacy in treating, preventing and/or managing arthritis(e.g., rheumatoid or psoriatic arthritis) can be assessed usingautoimmune animal models described, for example, in Williams et al.(2010) Chem Biol, 17(2):123-34, WO 2009/088986, WO2009/088880, and WO2011/008302; efficacy in treating, preventing and/or managing psoriasiscan be assessed using transgenic or knockout mouse model with targetedmutations in epidermis, vasculature or immune cells, mouse modelresulting from spontaneous mutations, and immuno-deficient mouse modelwith xenotransplantation of human skin or immune cells, all of which aredescribed, for example, in Boehncke et al. (2007) Clinics inDermatology, 25: 596-605; efficacy in treating, preventing and/ormanaging fibrosis or fibrotic condition can be assessed using theunilateral ureteral obstruction model of renal fibrosis (see Chevalieret al., Kidney International (2009) 75:1145-1152), the bleomycin inducedmodel of pulmonary fibrosis (see Moore and Hogaboam, Am. J. Physiol.Lung. Cell. Mol. Physiol. (2008) 294:L152-L160), a variety ofliver/biliary fibrosis models (see Chuang et al., Clin Liver Dis (2008)12:333-347 and Omenetti, A. et al. (2007) Laboratory Investigation87:499-514 (biliary duct-ligated model)), or a number of myelofibrosismouse models (see Varicchio, L. et al. (2009) Expert Rev. Hematol.2(3):315-334); efficacy in treating, preventing and/or managingscleroderma can be assessed using mouse model induced by repeated localinjections of bleomycin (“BLM”) described, for example, in Yamamoto etal. (1999) J Invest Dermatol 112: 456-462; efficacy in treating,preventing and/or managing dermatomyositis can be assessed usingmyositis mouse model induced by immunization with rabbit myosindescribed, for example, in Phyanagi et al. (2009) Arthritis &Rheumatism, 60(10): 3118-3127; efficacy in treating, preventing and/ormanaging lupus (e.g., CLE or SLE) can be assessed using various animalmodels described, for example, in Ghoreishi et al. (2009) Lupus, 19:1029-1035, Ohl et al. (2011) Journal of Biomedicine and Biotechnology,Article ID 432595 (14 pages), Xia et al. (2011) Rheumatology,50:2187-2196, Pau et al. (2012) PLoS ONE, 7(5):e36761 (15 pages),Mustafa et al. (2011) Toxicology, 290:156-168, Ichikawa et al. (2012)Arthritis and Rheumatism, 62(2): 493-503, Ouyang et al. (2012) J MolMed, DOI 10.1007/s00109-012-0866-3 (10 pages), Rankin et al. (2012)Journal of Immunology, 188:1656-1667; and efficacy in treating,preventing and/or managing Sjögren's syndrome can be assessed usingvarious mouse models described, for example, in Chiorini et al. (2009)Journal of Autoimmunity, 33: 190-196.

In one embodiment, provided herein is a method of treating, preventingand/or managing asthma. As used herein, “asthma” encompasses airwayconstriction regardless of the cause. Common triggers of asthma include,but are not limited to, exposure to an environmental stimulants (e.g.,allergens), cold air, warm air, perfume, moist air, exercise orexertion, and emotional stress. Also provided herein is a method oftreating, preventing and/or managing one or more symptoms associatedwith asthma. Examples of the symptoms include, but are not limited to,severe coughing, airway constriction and mucus production.

In one embodiment, provided herein is a method of treating, preventingand/or managing arthritis. As used herein, “arthritis” encompasses alltypes and manifestations of arthritis. Examples include, but are notlimited to, crystalline arthritis, osteoarthritis, psoriatic arthritis,gouty arthritis, reactive arthritis, rheumatoid arthritis and Reiter'sarthritis. In one embodiment, the disease or disorder is rheumatoidarthritis. In another embodiment, the disease or disorder is psoriaticarthritis. Also provided herein is a method of treating, preventingand/or managing one or more symptoms associated with arthritis. Examplesof the symptoms include, but are not limited to, joint pain, whichprogresses into joint deformation, or damages in body organs such as inblood vessels, heart, lungs, skin, and muscles.

In one embodiment, provided herein is a method of treating, preventingand/or managing psoriasis. As used herein, “psoriasis” encompasses alltypes and manifestations of psoriasis. Examples include, but are notlimited to, plaque psoriasis (e.g., chronic plaque psoriasis, moderateplaque psoriasis and severe plaque psoriasis), guttate psoriasis,inverse psoriasis, pustular psoriasis, pemphigus vulgaris, erythrodermicpsoriasis, psoriasis associated with inflammatory bowel disease (IBD),and psoriasis associated with rheumatoid arthritis (RA). Also providedherein is a method of treating, preventing and/or managing one or moresymptoms associated with psoriasis. Examples of the symptoms include,but are not limited to: red patches of skin covered with silvery scales;small scaling spots; dry, cracked skin that may bleed; itching; burning;soreness; thickened, pitted or ridged nails; and swollen and stiffjoints.

In one embodiment, provided herein is a method of treating, preventingand/or managing fibrosis and fibrotic condition. As used herein,“fibrosis” or “fibrotic condition encompasses all types andmanifestations of fibrosis or fibrotic condition. Examples include, butare not limited to, formation or deposition of tissue fibrosis; reducingthe size, cellularity (e.g., fibroblast or immune cell numbers),composition; or cellular content, of a fibrotic lesion; reducing thecollagen or hydroxyproline content, of a fibrotic lesion; reducingexpression or activity of a fibrogenic protein; reducing fibrosisassociated with an inflammatory response; decreasing weight lossassociated with fibrosis; or increasing survival.

In certain embodiments, the fibrotic condition is primary fibrosis. Inone embodiment, the fibrotic condition is idiopathic. In otherembodiments, the fibrotic condition is associated with (e.g., issecondary to) a disease (e.g., an infectious disease, an inflammatorydisease, an autoimmune disease, a malignant or cancerous disease, and/ora connective disease); a toxin; an insult (e.g., an environmental hazard(e.g., asbestos, coal dust, polycyclic aromatic hydrocarbons), cigarettesmoking, a wound); a medical treatment (e.g., surgical incision,chemotherapy or radiation), or a combination thereof.

In some embodiments, the fibrotic condition is associated with anautoimmune disease selected from scleroderma or lupus, e.g., systemiclupus erythematosus. In some embodiments, the fibrotic condition issystemic. In some embodiments, the fibrotic condition is systemicsclerosis (e.g., limited systemic sclerosis, diffuse systemic sclerosis,or systemic sclerosis sine scleroderma), nephrogenic systemic fibrosis,cystic fibrosis, chronic graft vs. host disease, or atherosclerosis.

In certain embodiments, the fibrotic condition is a fibrotic conditionof the lung, a fibrotic condition of the liver, a fibrotic condition ofthe heart or vasculature, a fibrotic condition of the kidney, a fibroticcondition of the skin, a fibrotic condition of the gastrointestinaltract, a fibrotic condition of the bone marrow or a hematopoietictissue, a fibrotic condition of the nervous system, a fibrotic conditionof the eye, or a combination thereof.

In other embodiment, the fibrotic condition affects a tissue chosen fromone or more of muscle, tendon, cartilage, skin (e.g., skin epidermis orendodermis), cardiac tissue, vascular tissue (e.g., artery, vein),pancreatic tissue, lung tissue, liver tissue, kidney tissue, uterinetissue, ovarian tissue, neural tissue, testicular tissue, peritonealtissue, colon, small intestine, biliary tract, gut, bone marrow,hematopoietic tissue, or eye (e.g., retinal) tissue.

In some embodiments, the fibrotic condition is a fibrotic condition ofthe eye. In some embodiments, the fibrotic condition is glaucoma,macular degeneration (e.g., age-related macular degeneration), macularedema (e.g., diabetic macular edema), retinopathy (e.g., diabeticretinopathy), or dry eye disease.

In certain embodiments, the fibrotic condition is a fibrotic conditionof the lung. In certain embodiments, the fibrotic condition of the lungis chosen from one or more of: pulmonary fibrosis, idiopathic pulmonaryfibrosis (IPF), usual interstitial pneumonitis (UIP), interstitial lungdisease, cryptogenic fibrosing alveolitis (CFA), bronchiectasis, andscleroderma lung disease. In one embodiment, the fibrosis of the lung issecondary to a disease, a toxin, an insult, a medical treatment, or acombination thereof. For example, the fibrosis of the lung can beassociated with (e.g., secondary to) one or more of: a disease processsuch as asbestosis and silicosis; an occupational hazard; anenvironmental pollutant; cigarette smoking; an autoimmune connectivetissue disorders (e.g., rheumatoid arthritis, scleroderma and systemiclupus erythematosus (SLE)); a connective tissue disorder such assarcoidosis; an infectious disease, e.g., infection, particularlychronic infection; a medical treatment, including but not limited to,radiation therapy, and drug therapy, e.g., chemotherapy (e.g., treatmentwith as bleomycin, methotrexate, amiodarone, busulfan, and/ornitrofurantoin). In one embodiment, the fibrotic condition of the lungtreated with the methods provided herein is associated with (e.g.,secondary to) a cancer treatment, e.g., treatment of a cancer (e.g.,squamous cell carcinoma, testicular cancer, Hodgkin's disease withbleomycin). In one embodiment, the fibrotic condition of the lung isassociated with an autoimmune connective tissue disorder (e.g.,scleroderma or lupus, e.g., SLE).

In certain embodiments, the fibrotic condition is a fibrotic conditionof the liver. In certain embodiments, the fibrotic condition of theliver is chosen from one or more of: fatty liver disease, steatosis(e.g., nonalcoholic steatohepatitis (NASH), cholestatic liver disease(e.g., primary biliary cirrhosis (PBC)), cirrhosis, alcohol inducedliver fibrosis, biliary duct injury, biliary fibrosis, orcholangiopathies. In other embodiments, hepatic or liver fibrosisincludes, but is not limited to, hepatic fibrosis associated withalcoholism, viral infection, e.g., hepatitis (e.g., hepatitis C, B orD), autoimmune hepatitis, non-alcoholic fatty liver disease (NAFLD),progressive massive fibrosis, exposure to toxins or irritants (e.g.,alcohol, pharmaceutical drugs and environmental toxins).

In certain embodiments, the fibrotic condition is a fibrotic conditionof the heart. In certain embodiments, the fibrotic condition of theheart is myocardial fibrosis (e.g., myocardial fibrosis associated withradiation myocarditis, a surgical procedure complication (e.g.,myocardial post-operative fibrosis), infectious diseases (e.g., Chagasdisease, bacterial, trichinosis or fungal myocarditis)); granulomatous,metabolic storage disorders (e.g., cardiomyopathy, hemochromatosis);developmental disorders (e.g., endocardial fibroelastosis);arteriosclerotic, or exposure to toxins or irritants (e.g., drug inducedcardiomyopathy, drug induced cardiotoxicity, alcoholic cardiomyopathy,cobalt poisoning or exposure). In certain embodiments, the myocardialfibrosis is associated with an inflammatory disorder of cardiac tissue(e.g., myocardial sarcoidosis). In some embodiments, the fibroticcondition is a fibrotic condition associated with a myocardialinfarction. In some embodiments, the fibrotic condition is a fibroticcondition associated with congestive heart failure.

In certain embodiments, the fibrotic condition is a fibrotic conditionof the kidney. In certain embodiments, the fibrotic condition of thekidney is chosen from one or more of: renal fibrosis (e.g., chronickidney fibrosis), nephropathies associated with injury/fibrosis (e.g.,chronic nephropathies associated with diabetes (e.g., diabeticnephropathy)), lupus, scleroderma of the kidney, glomerular nephritis,focal segmental glomerular sclerosis, IgA nephropathyrenal fibrosisassociated with human chronic kidney disease (CKD), chronic progressivenephropathy (CPN), tubulointerstitial fibrosis, ureteral obstruction,chronic uremia, chronic interstitial nephritis, radiation nephropathy,glomerulosclerosis, progressive glomerulonephrosis (PGN),endothelial/thrombotic microangiopathy injury, HIV-associatednephropathy, or fibrosis associated with exposure to a toxin, anirritant, or a chemotherapeutic agent. In one embodiment, the fibroticcondition of the kidney is scleroderma of the kidney. In someembodiments, the fibrotic condition of the kidney is transplantnephropathy, diabetic nephropathy, lupus nephritis, or focal segmentalglomerulosclerosis (FSGS).

In certain embodiments, the fibrotic condition is a fibrotic conditionof the skin. In certain embodiments, the fibrotic condition of the skinis chosen from one or more of: skin fibrosis (e.g., hypertrophicscarring, keloid), scleroderma, nephrogenic systemic fibrosis (e.g.,resulting after exposure to gadolinium (which is frequently used as acontrast substance for MRIs) in patients with severe kidney failure),and keloid.

In certain embodiments, the fibrotic condition is a fibrotic conditionof the gastrointestinal tract. In certain embodiments, the fibroticcondition is chosen from one or more of: fibrosis associated withscleroderma; radiation induced gut fibrosis; fibrosis associated with aforegut inflammatory disorder such as Barrett's esophagus and chronicgastritis, and/or fibrosis associated with a hindgut inflammatorydisorder, such as inflammatory bowel disease (IBD), ulcerative colitisand Crohn's disease. In some embodiments, the fibrotic condition of thegastrointestinal tract is fibrosis associated with scleroderma.

In certain embodiments, the fibrotic condition is a fibrotic conditionof the bone marrow or a hematopoietic tissue. In certain embodiments,the fibrotic condition of the bone marrow is an intrinsic feature of achronic myeloproliferative neoplasm of the bone marrow, such as primarymyelofibrosis (also referred to herein as agnogenic myeloid metaplasiaor chronic idiopathic myelofibrosis). In other embodiments, the bonemarrow fibrosis is associated with (e.g., is secondary to) a malignantcondition or a condition caused by a clonal proliferative disease. Inother embodiments, the bone marrow fibrosis is associated with ahematologic disorder (e.g., a hematologic disorder chosen from one ormore of polycythemia vera, essential thrombocythemia, myelodysplasia,hairy cell leukemia, lymphoma (e.g., Hodgkin or non-Hodgkin lymphoma),multiple myeloma or chronic myelogeneous leukemia (CML)). In yet otherembodiments, the bone marrow fibrosis is associated with (e.g.,secondary to) a non-hematologic disorder (e.g., a non-hematologicdisorder chosen from solid tumor metastasis to bone marrow, anautoimmune disorder (e.g., systemic lupus erythematosus, scleroderma,mixed connective tissue disorder, or polymyositis), an infection (e.g.,tuberculosis), or secondary hyperparathyroidism associated with vitaminD deficiency. In some embodiments, the fibrotic condition is idiopathicor drug-induced myelofibrosis. In some embodiments, the fibroticcondition of the bone marrow or hematopoietic tissue is associated withsystemic lupus erythematosus or scleroderma.

In one embodiment, provided herein is a method of treating, preventingand/or managing scleroderma. Scleroderma is a group of diseases thatinvolve hardening and tightening of the skin and/or other connectivetissues. Scleroderma may be localized (e.g., affecting only the skin) orsystemic (e.g., affecting other systems such as, e.g., blood vesselsand/or internal organs). Common symptoms of scleroderma includeRaynaud's phenomenon, gastroesophageal reflux disease, and skin changes(e.g., swollen fingers and hands, or thickened patches of skin). In someembodiments, the scleroderma is localized, e.g., morphea or linearscleroderma. In some embodiments, the condition is a systemic sclerosis,e.g., limited systemic sclerosis, diffuse systemic sclerosis, orsystemic sclerosis sine scleroderma.

Localized scleroderma (localized cutaneous fibrosis) includes morpheaand linear scleroderma. Morphea is typically characterized byoval-shaped thickened patches of skin that are white in the middle, witha purple border. Linear scleroderma is more common in children. Symptomsof linear scleroderma may appear mostly on one side of the body. Inlinear scleroderma, bands or streaks of hardened skin may develop on oneor both arms or legs or on the forehead. En coup de sabre (frontallinear scleroderma or morphea en coup de sabre) is a type of localizedscleroderma typically characterized by linear lesions of the scalp orface.

Systemic scleroderma (systemic sclerosis) includes, e.g., limitedsystemic sclerosis (also known as limited cutaneous systemic sclerosis,or CREST syndrome), diffuse systemic sclerosis (also known as diffusecutaneous systemic sclerosis), and systemic sclerosis sine scleroderma.CREST stands for the following complications that may accompany limitedscleroderma: calcinosis (e.g., of the digits), Raynaud's phenomenon,esophageal dysfunction, sclerodactyly, and telangiectasias. Typically,limited scleroderma involves cutaneous manifestations that mainly affectthe hands, arms, and face. Limited and diffuse subtypes aredistinguished based on the extent of skin involvement, with sparing ofthe proximal limbs and trunk in limited disease. See, e.g., Denton, C.P. et al. (2006), Nature Clinical Practice Rheumatology, 2(3):134-143.The limited subtype also typically involves a long previous history ofRaynaud's phenomenon, whereas in the diffuse subtype, onset of Raynaud'sphenomenon can be simultaneous with other manifestations or might occurlater. Both limited and diffuse subtypes may involve internal organs.Typical visceral manifestations of limited systemic sclerosis includeisolated pulmonary hypertension, severe bowel involvement, and pulmonaryfibrosis. Typical visceral manifestations of diffuse systemic sclerosisinclude renal crisis, lung fibrosis, and cardiac disease. Diffusesystemic sclerosis typically progresses rapidly and affects a large areaof the skin and one or more internal organs (e.g., kidneys, esophagus,heart, or lungs). Systemic sclerosis sine scleroderma is a rare disorderin which patients develop vascular and fibrotic damage to internalorgans in the absence of cutaneous sclerosis.

In one embodiment, provided herein is a method of treating, preventingand/or managing inflammatory myopathies. As used herein, “inflammatorymyopathies” encompass all types and manifestations of inflammatorymyopathies. Examples include, but are not limited to, muscle weakness(e.g., proximal muscle weakness), skin rash, fatigue after walking orstanding, tripping or falling, dysphagia, dysphonia, difficultybreathing, muscle pain, tender muscles, weight loss, low-grade fever,inflamed lungs, light sensitivity, calcium deposits (calcinosis) underthe skin or in the muscle, as well as biological concomitants ofinflammatory myopathies as disclosed herein or as known in the art.Biological concomitants of inflammatory myopathies (e.g.,dermatomyositis) include, e.g., altered (e.g., increased) levels ofcytokines (e.g., Type I interferons (e.g., IFN-α and/or IFN-β),interleukins (e.g., IL-6, IL-10, IL-15, IL-17 and IL-18), and TNF-α),TGF-β, B-cell activating factor (BAFF), overexpression of IFN induciblegenes (e.g., Type I IFN inducible genes). Other biological concomitantsof inflammatory myopathies can include, e.g., an increased erythrocytesedimentation rate (ESR) and/or elevated level of creatine kinase.Further biological concomitants of inflammatory myopathies can includeautoantibodies, e.g., anti-synthetase autoantibodies (e.g., anti-Jo1antibodies), anti-signal recognition particle antibodies (anti-SRP),anti-Mi-2 antibodies, anti-p155 antibodies, anti-PM/Sci antibodies, andanti-RNP antibodies.

The inflammatory myopathy can be an acute inflammatory myopathy or achronic inflammatory myopathy. In some embodiments, the inflammatorymyopathy is a chronic inflammatory myopathy (e.g., dermatomyositis,polymyositis, or inclusion body myositis). In some embodiments, theinflammatory myopathy is caused by an allergic reaction, another disease(e.g., cancer or a connective tissue disease), exposure to a toxicsubstance, a medicine, or an infectious agent (e.g., a virus). In someembodiments, the inflammatory myopathy is associated with lupus,rheumatoid arthritis, or systemic sclerosis. In some embodiments, theinflammatory myopathy is idiopathic. In some embodiments, theinflammatory myopathy is selected from polymyositis, dermatomyositis,inclusion body myositis, and immune-mediated necrotizing myopathy. Insome embodiments, the inflammatory myopathy is dermatomyositis.

In another embodiment, provided herein is a method of treating,preventing and/or managing a skin condition (e.g., a dermatitis). Insome embodiments, the methods provided herein can reduce symptomsassociated with a skin condition (e.g., itchiness and/or inflammation).In some such embodiments, the compound provided herein is administeredtopically (e.g., as a topical cream, eye-drop, nose drop or nasalspray). In some such embodiments, the compound is a PI3K-delta inhibitor(e.g., a PI3K inhibitor that demonstrates greater inhibition ofPI3K-delta than of other PI3K isoforms). In some embodiments, thePI3K-delta inhibitor prevents mast cell degranulation.

As used herein, “skin condition” includes any inflammatory condition ofthe skin (e.g., eczema or dermatitis, e.g., contact dermatitis, atopicdermatitis, dermatitis herpetiformis, seborrheic dermatitis, nummulardermatitis, stasis dermatitis, perioral dermatitis), as well asaccompanying symptoms (e.g., skin rash, itchiness (pruritis), swelling(edema), hay fever, anaphalaxis). Frequently, such skin conditions arecaused by an allergen. As used herein, a “skin condition” also includes,e.g., skin rashes (e.g., allergic rashes, e.g., rashes resulting fromexposure to allergens such as poison ivy, poison oak, or poison sumac,or rashes caused by other diseases or conditions), insect bites, minorburns, sunburn, minor cuts, and scrapes. In some embodiments, thesymptom associated with inflammatory myopathy, or the skin condition orsymptom associated with the skin condition, is a skin rash or itchiness(pruritis) caused by a skin rash.

The skin condition (e.g., the skin rash) may be spontaneous, or it maybe induced, e.g., by exposure to an allergen (e.g., poison ivy, poisonoak, or poison sumac), drugs, food, insect bite, inhalants, emotionalstress, exposure to heat, exposure to cold, or exercise. In someembodiments, the skin condition is a skin rash (e.g., a pruritic rash,e.g., utricaria). In some embodiments, the skin condition is an insectbite. In some embodiments, the skin condition is associated with anotherdisease (e.g., an inflammatory myopathy, e.g., dermatomyositis).

In some embodiments, the subject (e.g., the subject in need of treatmentfor an inflammatory myopathy and/or a skin condition) exhibits anelevated level or elevated activity of IFN-α, TNF-α, IL-6, IL-8, IL-1,or a combination thereof. In certain embodiments, the subject exhibitsan elevated level of IFN-α. In some embodiments, treating (e.g.,decreasing or inhibiting) the inflammatory myopathy, or the skincondition, comprises inhibiting (e.g., decreasing a level of, ordecreasing a biological activity of) one or more of IFN-α, TNF-α, IL-6,IL-8, or IL-1 in the subject or in a sample derived from the subject. Insome embodiments, the method decreases a level of IFN-α, TNF-α, IL-6,IL-8, or IL-1 in the subject or in a sample derived from the subject. Insome embodiments, the method decreases a level of IFN-α in the subjector in a sample derived from the subject. In some embodiments, the levelof IFN-α, TNF-α, IL-6, IL-8, or IL-1 is the level assessed in a sampleof whole blood or PBMCs. In some embodiments, the level of IFN-α, TNF-α,IL-6, IL-8, or IL-1 is the level assessed in a sample obtained by a skinbiopsy or a muscle biopsy. In some embodiments, the sample is obtainedby a skin biopsy.

In one embodiment, provided herein is a method of treating, preventingand/or managing myositis. As used herein, “myositis” encompasses alltypes and manifestations of myositis. Examples include, but are notlimited to, myositis ossificans, fibromyositis, idiopathic inflammatorymyopathies, dermatomyositis, juvenile dermatomyositis, polymyositis,inclusion body myositis and pyomyositis. In one embodiment, the diseaseor disorder is dermatomyositis. Also provided herein is a method oftreating, preventing and/or managing one or more symptoms associatedwith myositis. Examples of the symptoms include, but are not limited to:muscle weakness; trouble lifting arms; trouble swallowing or breathing;muscle pain; muscle tenderness; fatigue; fever; lung problems;gastrointestinal ulcers; intestinal perforations; calcinosis under theskin; soreness; arthritis; weight loss; and rashes.

In one embodiment, provided herein is a method of treating, preventingand/or managing lupus. As used herein, “lupus” refers to all types andmanifestations of lupus. Examples include, but are not limited to,systemic lupus erythematosus; lupus nephritis; cutaneous manifestations(e.g., manifestations seen in cutaneous lupus erythematosus, e.g., askin lesion or rash); CNS lupus; cardiovascular, pulmonary, hepatic,hematological, gastrointestinal and musculoskeletal manifestations;neonatal lupus erythematosus; childhood systemic lupus erythematosus;drug-induced lupus erythematosus; anti-phospholipid syndrome; andcomplement deficiency syndromes resulting in lupus manifestations. Inone embodiment, the lupus is systemic lupus erythematosus (SLE),cutaneous lupus erythematosus (CLE), drug-induced lupus, or neonatallupus. In another embodiment, the lupus is a CLE, e.g., acute cutaneouslupus erythematosus (ACLE), subacute cutaneous lupus erythematosus(SCLE), intermittent cutaneous lupus erythematosus (also known as lupuserythematosus tumidus (LET)), or chronic cutaneous lupus. In someembodiments, the intermittent CLE is chronic discloid lupuserythematosus (CDLE) or lupus erythematosus profundus (LEP) (also knownas lupus erythematosus panniculitis). Types, symptoms, and pathogenesisof CLE are described, for example, in Wenzel et al. (2010), Lupus, 19,1020-1028.

In one embodiment, provided herein is a method of treating, preventingand/or managing Sjögren's syndrome. As used herein, “Sjögren's syndrome”refers to all types and manifestations of Sjögren's syndrome. Examplesinclude, but are not limited to, primary and secondary Sjögren'ssyndrome. Also provided herein is a method of treating, preventingand/or managing one or more symptoms associated with Sjögren's syndrome.Examples of the symptoms include, but are not limited to: dry eyes; drymouth; joint pain; swelling; stiffness; swollen salivary glands; skinrashes; dry skin; vaginal dryness; persistent dry cough; and prolongedfatigue.

In some embodiments, provided herein is a method of treating a bonedisorder in a subject that comprises administering to said subject atherapeutically effective amount of a compound provided herein (e.g., aPI3K-γ selective compound provided herein), or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein. As usedherein, “bone disorder” encompasses all types and manifestations of bonedisorders. Exemplary bone disorders include, but are not limited to,bone cancer, bone metastases, osteoporosis, fracture repair, avascularnecrosis (osteonecrosis), bone spur (osteophytes), craniosynostosis,Coffin-Lowry syndrome, fibrodysplasia ossificans progressive, fibrousdysplasia, Fong Disease (Nail-patella syndrome), Giant cell tumor ofbone, Greenstick Fracture, hypophosphatasia, Klippel-Feil syndrome,metabolic bone disease, osteoarthritis, osteitis deformans (Paget'sdisease of bone), osteitis fibrosa cystica (osteitis fibrosa or VonRecklinghausen's disease of bone), osteitis pubis, condensing osteitis(osteitis condensas), osteochondritis dissecans, osteochondroma (bonetumor), osteogenesis imperfect, osteomalacia, osteomyelitis, osteopenia,osteopetrosis, porotic hyperostosis, primary hyperparathyroidism, renalosteodystrophy, Salter-Harris fractures, and water on the knee. In oneembodiment, the bone disorder is a systemical bone disorder. In anotherembodiment, the bone disorder is a topical bone disorder. In oneembodiment, the bone disorder is associated with excess bone formation.In another embodiment, the bone disorder is associated with excess boneresorption. In one embodiment, without being limited by a particulartheory, a compound provided herein inhibits differentiation ofosteoclasts from bone marrow macrophages.

In some embodiments, a symptom associated with the disease or disorderprovided herein is reduced by at least 10%, at least 20%, at least 30%,at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, or at least 95% relative to a control level. The controllevel includes any appropriate control as known in the art. For example,the control level can be the pre-treatment level in the sample orsubject treated, or it can be the level in a control population (e.g.,the level in subjects who do not have the disease or disorder or thelevel in samples derived from subjects who do not have the disease ordisorder). In some embodiments, the decrease is statisticallysignificant, for example, as assessed using an appropriate parametric ornon-parametric statistical comparison.

Methods of Treatment, Prevention and/or Management for Pulmonary orRespiratory Disorders

Without being limited by a particular theory, it was found thatadministering a compound provided herein (e.g., Compound 1) byinhalation can accord various therapeutic benefits as described hereinin treating, preventing and/or managing pulmonary or respiratorydiseases. Accordingly, in certain embodiments, provided herein is amethod of treating, preventing, and/or managing a pulmonary orrespiratory disease in a subject, comprising administering to a subjectin need thereof by inhalation a therapeutically or prophylacticallyeffective amount of a compound provided herein, or an enantiomer, amixture of enantiomers, or a mixture of two or more diastereomersthereof, or a pharmaceutically acceptable form thereof.

In addition, without being limited by a particular theory, it was foundthat administering a compound provided herein by inhalation results in aprolonged retainment of the compound in patient's lung. Thus, in someembodiments, provided herein is a method of eliciting prolongedanti-inflammatory effect in lung in a subject suffering from a pulmonaryor respiratory disease, comprising administering to the subject byinhalation a therapeutically or prophylactically effective amount of acompound provided herein, or an enantiomer, a mixture of enantiomers, ora mixture of two or more diastereomers thereof, or a pharmaceuticallyacceptable form thereof, wherein the compound is retained in lung for aprolonged period (e.g., a period longer than what is provided by oraladministration).

In some embodiments, the compound is retained in lung for about hour,about 3 hours, about 6 hours, about 12 hours, about 24 hours, about 48hours, or about 72 hours longer than what is provided by oraladministration.

In some embodiments, more than 80%, more than 70%, more than 60%, morethan 50%, more than 40%, more than 30%, or more than 20% of the amountof the compound as initially administered to patient remains in lung at24 hours after administration by inhalation.

In some embodiments, the concentration of the compound in lung followingadministration by inhalation is about 100, about 200, about 500, about1000, about 2000, about 3000, about 4000, about 5000, about 6000, about7000, about 8000, about 9000, or about 10000 times higher than theplasma concentration of the compound at about 5 hours after theadministration. In some embodiments, the concentration of the compoundin lung following administration by inhalation is about 100, about 200,about 500, about 1000, about 2000, about 3000, about 4000, about 5000,about 6000, about 7000, about 8000, about 9000, or about 10000 timeshigher than the plasma concentration of the compound at about 12 hoursafter the administration. In some embodiments, the concentration of thecompound in lung following administration by inhalation is about 100,about 200, about 500, about 1000, about 2000, about 3000, about 4000,about 5000, about 6000, about 7000, about 8000, about 9000, or about10000 times higher than the plasma concentration of the compound atabout 24 hours after the administration.

In some embodiments, the compound is administered at a dose of less than0.01 μg/kg/day, less than 0.02 μg/kg/day, less than 0.05 μg/kg/day, lessthan 0.1 μg/kg/day, less than 0.2 μg/kg/day, less than 0.5 μg/kg/day,less than 1 μg/kg/day, less than 2 μg/kg/day, less than 5 μg/kg/day,less than 10 μg/kg/day, less than 20 μg/kg/day, less than 50 μg/kg/day,or less than 100 μg/kg/day. In some embodiments, the compound isadministered at a dose of about 0.01 μg/kg/day, about 0.02 μg/kg/day,about 0.05 μg/kg/day, about 0.1 μg/kg/day, about 0.2 μg/kg/day, about0.5 μg/kg/day, about 1 μg/kg/day, about 2 μg/kg/day, about 5 μg/kg/day,about 10 μg/kg/day, about 20 μg/kg/day, about 50 μg/kg/day, or about 100μg/kg/day. In some embodiments, the compound is administered at a doseof from about 0.01 μg/kg/day to about 100 μg/kg/day, from about 0.01μg/kg/day to about 50 μg/kg/day, from about 0.01 μg/kg/day to about 20μg/kg/day, from about 0.01 μg/kg/day to about 10 μg/kg/day, from about0.01 μg/kg/day to about 5 μg/kg/day, from about 0.01 μg/kg/day to about1 μg/kg/day, from about 0.05 μg/kg/day to about 1 μg/kg/day, or fromabout 0.1 μg/kg/day to about 1 μg/kg/day.

In one embodiment, the compound is administered once daily (QD). Inanother embodiment, the compound is administered twice daily (BID). Inanother embodiment, the compound is administered three time daily (TID).In another embodiment, the compound is administered four times daily(QID).

In one embodiment, the subject is a mammal. In another embodiment, thesubject is a human.

In one embodiment, provided herein is a method of treating, preventing,and/or managing a pulmonary or respiratory disease in a subject,comprising administering to a subject in need thereof by inhalation atherapeutically or prophylactically effective amount of a PI3Kγinhibitor, or an enantiomer, a mixture of enantiomers, or a mixture oftwo or more diastereomers thereof, or a pharmaceutically acceptable formthereof. In some embodiments, the PI3Kγ inhibitor has a delta/gammaselectivity ratio of greater than about 1 to <10, greater than about 10to <50, or greater than about 50 to <350. In some embodiments, the PI3Kγinhibitor has a delta/gamma selectivity ratio of greater than about 1,greater than about 5, greater than about 10, greater than about 15,greater than about 20, greater than about 25, greater than about 50,greater than about 75, greater than about 100, greater than about 150,greater than about 200, greater than about 250, greater than about 300,greater than about 350, greater than about 500, or greater than about1000.

In one embodiment, provided herein is a method of treating, preventing,and/or managing a pulmonary or respiratory disease in a subject,comprising administering to a subject in need thereof by inhalation atherapeutically or prophylactically effective amount of Compound 1, oran enantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof. Inone embodiment, Compound 1 is a PI3Kγ inhibitor. In one embodiment, thecompound, e.g., Compound 1 has a delta/gamma selectivity ratio ofgreater than about 1 to <10, greater than about 10 to <50, or greaterthan about 50 to <350. In one embodiment, the compound, e.g., Compound 1has a delta/gamma selectivity ratio of greater than about 50 to <350. Inone embodiment, the compound, e.g., Compound 1 has a delta/gammaselectivity ratio of greater than about 50 to <150. In one embodiment,the compound, e.g., Compound 1 has a delta/gamma selectivity ratio ofgreater than about 75 to <125. In one embodiment, the compound has adelta/gamma selectivity ratio of greater than about 100. In someembodiments, the compound has a delta/gamma selectivity ratio of greaterthan about 1, greater than about 5, greater than about 10, greater thanabout 15, greater than about 20, greater than about 25, greater thanabout 50, greater than about 75, greater than about 100, greater thanabout 150, greater than about 200, greater than about 250, greater thanabout 300, greater than about 350, greater than about 500, or greaterthan about 1000.

Administration by Inhalation

Many diseases of the respiratory tract are known to respond to treatmentby the direct application of therapeutic agents by inhalation. Suchadministration can result in the better utilization of the medicament inthat the drug is deposited directly at the desired site and where itsaction may be required. Therefore, without being limited by a particulartheory, administration by inhalation can significantly reduce the doserequired to achieve therapeutic efficacy, which, in turn can result inmarked reduction of undesired side effects and cost of medicament. It istypically accepted in the industry that the bioavailability of the drugis optimum when the drug particles delivered to the respiratory tractare between 1 to 5 microns in size.

Various methods and devices can be used to deliver a compound providedherein by inhalation. The inhalable formulation can be administered viathe mouth or nose ultimately for pulmonary delivery thereof. Forexample, dry powder inhalers (DPIs), which usually have a means forintroducing the drug (active drug plus carrier) into a high velocity airstream, can be used to practice the methods provided herein. The highvelocity air stream is used as the primary mechanism for breaking up thecluster of micronized particles or separating the drug particles fromthe carrier. Inhalation devices useful for dispensing powder forms ofmedicament such as those described in U.S. Pat. Nos. 3,507,277;3,518,992; 3,635,219; 3,795,244; and 3,807,400, are encompassed by thecurrent disclosure. In certain embodiments, such devices also includepropeller means, which upon inhalation aid in dispensing the powder outof the capsule, so that it is not necessary to rely solely on theinhaled air to suction powder from the capsule. (See, e.g., U.S. Pat.Nos. 2,517,482; 3,831,606; 3,948,264; and 5,458, 135, all of which areincorporated herein by reference). In certain embodiments, utilizationof vibration to facilitate suspension of power into an inhaled gasstream and which utilizes synthetic jetting to aerosolize drug powderfrom a blister pack is also provided herein. (See, e.g., U.S. Pat. Nos.7,318,434 and 7,334,577, incorporated herein by reference). In someembodiments, controlled aliquots or doses of a medication orpre-packaged drug in a blister pack, which includes a frangible crownedtop element which can be conical, conical with a rounded point, rounded,such as those described in U.S. Pat. No. 7,080,644, are alsoencompassed.

In certain embodiments, a compound provided herein is administered usingmetered dose inhalers (MDIs). MDIs typically have a pressurized canisterfilled with a liquid propellant. The drug is either suspended ordissolved in the propellant. The MDIs have a metering valve for meteringout a known quantity of the propellant and hence the drug. When thecanister is depressed against the MDI housing a known quantity of thepropellant is discharged. The propellant evaporates leaving behind afine aerosol of the drug suitable for inhalation by the patient. Incertain embodiments, MDIs that contain a breath actuation mechanism aspacer are also encompassed herein.

In some embodiments, a compound provided herein is administered usingnebulizers, such as the jet nebulizers. Nebulizers produce a fineaerosol mist/droplets which carry the drug either as a suspension ordissolved in the aqueous medium. The jet nebulizers use compressed airto atomize the aqueous solution. A drug can be administered to a patientwith repetitive non-forced inhalation over a prolonged period of time.

Examples of devices suitable for such pulmonary delivery include, butare not limited to, air-jet, ultrasonic, or vibrating-mesh devices suchas Pari LC Star, Aeroeclipse II, Prodose (HaloLite), Acorn II, TUp-draft II, Sidestream, AeroTech II, Mini heart, MisterNeb, Sonix 2000,MABISMist II and other suitable aerosol systems. In some embodiments,the nebulizer is a vibrating-mesh nebulizer that could include anAERONEB PRO, AERONEB SOLO, AERONEB GO, AERONEB LAB, OMRON MICROAIR, PARTEFLOW, RESPIRONICS I-NEB, or other suitable devices.

Pulmonary or Respiratory Diseases

Provided herein is a method of treating, preventing, and/or managingpulmonary or respiratory disease using a compound provided herein.Examples of pulmonary or respiratory disease include, but are notlimited to, lung inflammation, chronic obstructive pulmonary disease,asthma, pneumonia, hypersensitivity pneumonitis, pulmonary infiltratewith eosinophilia, environmental lung disease, pneumonia,bronchiectasis, cystic fibrosis, interstitial lung disease, postinflammatory pulmonary fibrosis, primary pulmonary hypertension,pulmonary thromboembolism, disorders of the pleura, disorders of themediastinum, disorders of the diaphragm, disorders of the larynx,disorders of the trachea, acute lung injury, hypoventilation,hyperventilation, sleep apnea, acute respiratory distress syndrome,mesothelioma, sarcoma, graft rejection, graft versus host disease, lungcancer, allergic rhinitis, allergy, allergic bronchopulmonaryaspergillosis, asbestosis, aspergilloma, aspergillosis, bronchiectasis,chronic bronchitis, emphysema, eosinophilic pneumonia, idiopathicpulmonary fibrosis, idiopathic interstitial pneumonia, non-specificinterstitial pneumonia (NSIP), bronchiolitis obliterans with organizingpneumonia (BOOP, also called cryptogenic organizing pneumonia or COP),lymphocytic interstitial pneumonia (LIP), acute interstitial pneumonitisinvasive pneumococcal disease, pneumococcal pneumonia, influenza,nontuberculous mycobacteria, pleural effusion, a pleural cavity disease,empyema, pleurisy, pneumoconiosis, pneumocytosis, respiratory viralinfection, acute bronchitis, aspiration pneumonia, ventilator-associatedpneumonia, pneumocystic jiroveci pneumonia, pneumonia, pulmonaryactinomycosis, pulmonary alveolar proteinosis, pulmonary anthrax,pulmonary edema, pulmonary embolus, pulmonary embolism, acute chestsyndrome, idiopathic pulmonary hemosiderosis, pulmonary hemorrhage,pulmonary hyperplasia, pulmonary inflammation, pulmonary histiocytosisX, eosinophilic granuloma, pulmonary Langerhan's cell histiocytosis,occupational lung disease, pneumopathy due to inhalation of dust,respiratory conditions due to chemical fumes and vapors, lipoidpneumonia, pulmonary hypertension, pulmonary arterial hypertension,pulmonary nocardiosis, pulmonary tuberculosis, pulmonary veno-occlusivedisease, pulmonary vascular disease, rheumatoid lung disease, connectivetissue disease-associated interstitial lung disease (e.g., systemicsclerosis (SSc or scleroderma)-associated interstitial lung disease,polymyositis-associated interstitial lung disease,dermatomyositis-associated interstitial lung disease, rheumatoidarthritis-associated interstitial lung disease, systemic lupuserythematosus-associated interstitial lung disease, interstitial lungdisease associated with Sjögren's syndrome, mixed connective tissuedisease-associated interstitial lung disease, and ankylosingspondylitis-associated interstitial lung disease), a restrictive lungdisease, a respiratory tract infection (upper and lower), sarcoidosis,Wegener's granulomatosis (also known as granulomatosis with polyangiitis(GPA) or necrotizing granulomatous vasculitis (NGV)), Churg-StraussSyndrome, microscopic polyangiitis (MPA), small cell lung carcinoma,non-small cell lung carcinoma, lymphangioleiomyomatosis (LAM),radiation-induced lung disease (also known as radiation pneumonitis),pulmonary vasculitis, viral pneumonia, pneumococcal pneumonia, bacterialpneumonia, bronchopneumonia, epithelial tumors, papillomas, adenomas,squamous cell carcinoma, small cell carcinoma, adenocarcinoma, largecell carcinoma, adenosquamous carcinoma, carcinoid tumor, carcinoma ofsalivary-gland type, soft tissue tumors, localized fibrous tumor,epithelioid hemangioendothelioma, pleuropulmonary blastoma, chondroma,calcifying fibrous pseudotumor of the pleura, congenital peribronchialmyofibroblastic tumor, diffuse pulmonary lymphangiomatosis, desmoplasticsmall round cell tumor, mesothelial tumors, adenomatoid tumor,epithelioid mesothelioma, sarcomatoid mesothelioma, biphasicmesothelioma, hamartoma, sclerosing hemangioma, clear cell tumor, germcell neoplasms, thymona, melanoma, and secondary tumor. In certainembodiments, provided herein is a method of treating, preventing, and/ormanaging a lymphoproliferative disease using a compound provided herein.Examples of lymphoproliferative disease include, but are not limited to,lymphoid interstitial pneumonia, nodular lymphoid hyperplasia, andlymphomatoid granulomatosis.

In certain embodiments, the pulmonary or respiratory disease to betreated, prevented and/or managed using a compound provided herein is anobstructive lung disease or disorder. In some embodiments, theobstructive lung disease is acute respiratory distress syndrome (ARDS),asthma, bronchiectasis, bronchiolectasis, bronchiolitis, bronchitis,chronic obstructive pulmonary disease (COPD), or emphysema.

Chronic Obstructive Pulmonary Disease

In one embodiment, said obstructive lung disease or disorder is chronicobstructive pulmonary disease (COPD), e.g., as diagnosed by a forcedexpiratory air volume in 1 second (FEV₁) to forced vital capacity (FVC)ratio of less than 0.7. In another embodiment, administration of acompound provided herein results in a detectable rise in the FEV₁/FECratio above 0.7 after administration, e.g., a rise of 0.02, 0.03, 0.04,0.05, 0.06, 0.07, 0.08, 0.09, 0.10, or more or more.

In one embodiment, provided herein is a method of reducing a COPDassociated symptom in a subject, comprising administering a compoundprovided herein (e.g., Compound 1), or an enantiomer, a mixture ofenantiomers, or a mixture of two or more diastereomers thereof, or apharmaceutically acceptable form thereof, by inhalation in an amountsufficient to reduce the COPD associated symptom. In one embodiment, thesubject is a mammalian subject, e.g., an animal model or as part oftherapeutic protocol. In one embodiment, the compound is used as asingle agent or in combination with another agent or therapeuticmodality.

In one embodiment, provided herein is a method of treating, preventing,and/or managing COPD in a subject, comprising administering an effectiveamount of a compound provided herein (e.g., Compound 1), or anenantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof, toa subject in need thereof by inhalation. In one embodiment, the compoundis administered as a single agent. In another embodiment, the compoundis administered in combination with another agent or therapeuticmodality.

As used herein, and unless otherwise specified, “COPD” or a “symptom”associated with COPD encompasses all types of manifestation of COPD asdisclosed herein or as known in the art. Examples of COPD include, butare not limited to, emphysema, chronic bronchitis, and bronchiectasis.Examples of symptom of COPD include, but are not limited to, wheezing,coughing, chest tightness, shortness of breath, difficulty in breathing,coughing up mucus/phlegm, and use of accessory muscle. Symptoms areoften worse at night or in the early morning, or in response to exerciseor cold air. In one embodiment, the symptom of asthma is shortness ofbreath or difficulty in breathing.

As used herein, and unless otherwise specified, to “decrease,”“ameliorate,” “reduce,” “inhibit,” “treat” (or the like) COPD or asymptom associated with COPD includes reducing the severity and/orfrequency of one or more symptoms of COPD, as well as preventing COPDand/or one or more symptoms of COPD (e.g., by reducing the severityand/or frequency of flares of symptoms).

In some embodiments, the symptom is reduced by at least about 2%, atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 40%, atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, or at least about 95% relative to a controllevel. The control level includes any appropriate control as known inthe art. For example, the control level can be the pre-treatment levelin the sample or subject treated, or it can be the level in a controlpopulation (e.g., the level in subjects who do not have COPD or thelevel in samples derived from subjects who do not have COPD). In someembodiments, the decrease is statistically significant, for example, asassessed using an appropriate parametric or non-parametric statisticalcomparison.

In certain embodiments, the subject is an animal model of COPD, a humanwith COPD, or a subject (e.g., a human) at risk for developing COPD. Insome embodiments, the subject is a human who has a family history ofCOPD, who carries a gene associated with COPD, who is positive for abiomarker associated with COPD, or a combination thereof. In someembodiments, the subject has been diagnosed with COPD. In someembodiments, the subject has one or more signs or symptoms associatedwith COPD. In some embodiments, the subject is at risk for developingCOPD (e.g., the subject carries a gene that, individually, or incombination with other genes or environmental factors, is associatedwith development of COPD).

In one embodiment, the subject has been previously diagnosed of COPD orhas episodic symptoms of airflow obstruction (e.g., shortness of breath,wheezing and/or chest tightness) for at least 1 week, 2 weeks, 1 month,2 months, 3 months, 6 months, 9 months, 12 months before a compoundprovided herein (e.g., Compound 1), or an enantiomer, a mixture ofenantiomers, or a mixture of two or more diastereomers thereof, or apharmaceutically acceptable form thereof, is administered. In oneembodiment, the subject has been previously diagnosed of COPD or hasepisodic symptoms of airflow obstruction (e.g., wheezing and/or chesttightness) for at least 6 months before a compound provided herein(e.g., Compound 1), or an enantiomer or a mixture of enantiomersthereof, or a pharmaceutically acceptable salt, solvate, hydrate,co-crystal, clathrate, or polymorph thereof, is administered.

In some embodiments, the subject has been previously treated for COPD.In some embodiments, the subject has been previously treated for COPDbut are non-responsive to standard therapies. In one embodiment, thestandard therapy is steroid, e.g., corticosteroids. In some embodiments,the subject has developed steroid resistance, e.g., from previoustreatment with steroids. In some embodiments, the subject can haveinherent steroid resistance that is not a result of previous treatments.Steroid resistance can be overcome by a PI3K inhibitor, e.g., a compoundprovided herein (e.g., Compound 1). Thus, combination therapy with acompound provided herein and a steroid can be beneficial. In oneembodiment, provided herein is a method of treating, preventing, and/ormanaging COPD in a subject, comprising administering an effective amountof a compound provided herein (e.g., Compound 1), or an enantiomer, amixture of enantiomers, or a mixture of two or more diastereomersthereof, or a pharmaceutically acceptable form thereof, to a subject inneed thereof, wherein the subject has been previously administered atherapy for COPD.

In some embodiments, the subject has not been previously treated forCOPD.

In one embodiment, without being limited by any particular theory,administering an effective amount of a compound provided herein (e.g.,Compound 1), or an enantiomer, a mixture of enantiomers, or a mixture oftwo or more diastereomers thereof, or a pharmaceutically acceptable formthereof, does not result in, or results in reduced, one or more commonside effects of COPD treatment. The common side effects of COPDtreatment include, but are not limited to: allergic reactions such asrashes, hives, swelling of the face, mouth and tongue, and breathingproblems; sudden breathing problems; effects on heart such as increasedblood pressure, fast and irregular heartbeat, and chest pain; effects onnervous system such as tremor and nervousness; reduced adrenal function;changes in blood contents; weakened immune system and higher chance ofinfections; lower bone mineral density; eye problems such as glaucomaand cataracts; slowed growth in children; pneumonia; thrush in the mouthand throat; throat irritation; hoarseness and voice changes; viralrespiratory infections; headache; and muscle and bone pain.

In some embodiments, the side effect is reduced by at least about 2%, atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 40%, atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, or at least about 95% relative to a controllevel. The control level includes any appropriate control as known inthe art. For example, the control level can be the side effect level inthe subject treated with other COPD therapies (e.g., albuterol,levalbuterol, ipratropium, tiotropium, terbutaline, theophylline,formoterol, salmeterol, flucatisone, methylprednisone, and prednisone).In some embodiments, the decrease is statistically significant, forexample, as assessed using an appropriate parametric or non-parametricstatistical comparison.

Asthma

In another specific embodiment, said obstructive lung disease ordisorder is asthma. In some embodiments, administration of a compoundprovided herein results in a detectable improvement in one or moresymptoms of asthma, e.g., airway obstruction, as determined byspirometry or a peak flow meter.

In one embodiment, provided herein is a method of reducing an asthmaassociated symptom in a subject, comprising administering a compoundprovided herein (e.g., Compound 1), or an enantiomer, a mixture ofenantiomers, or a mixture of two or more diastereomers thereof, or apharmaceutically acceptable form thereof, by inhalation in an amountsufficient to reduce the asthma associated symptom. In one embodiment,the subject is a mammalian subject, e.g., an animal model or as part oftherapeutic protocol. In one embodiment, the compound is used as asingle agent or in combination with another agent or therapeuticmodality.

In one embodiment, provided herein is a method of treating, preventing,and/or managing asthma in a subject, comprising administering aneffective amount of a compound provided herein (e.g., Compound 1), or anenantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof toa subject in need thereof by inhalation. In one embodiment, the compoundis administered as a single agent. In another embodiment, the compoundis administered in combination with another agent or therapeuticmodality.

As used herein, and unless otherwise specified, “asthma” or a “symptom”associated with asthma encompasses all types of manifestation of asthmaas disclosed herein or as known in the art. Examples of asthma include,but are not limited to, severe and/or refractory asthma, atopic(extrinsic) asthma, non-atopic (intrinsic) asthma, type 1 brittleasthma, type 2 brittle asthma, asthma attack, status asthmaticus,exercise-induced asthma, or occupational asthma. In one embodiment, theasthma is severe or refractory asthma. Examples of symptom of asthmainclude, but are not limited to, wheezing, coughing, chest tightness,shortness of breath, and use of accessory muscle. Symptoms are oftenworse at night or in the early morning, or in response to exercise orcold air. Asthma is clinically classified according to the frequency ofsymptoms, forced expiratory volume in 1 second (FEV₁), and peakexpiratory flow rate. In one embodiment, the symptom of asthma iswheezing or chest tightness.

As used herein, and unless otherwise specified, “asthma” or a “symptom”associated with asthma also encompasses biological concomitants ofasthma as disclosed herein or as known in the art. Examples include, butare not limited to, immune complexes, elevated levels of cytokines(e.g., interferons (e.g., Type I interferons, e.g., IFN-α and/or IFN-β);interleukins (e.g., IL-6, IL-8, IL-1, and IL-18) and TNF-α), elevatedlevels of anti-dsDNA autoantibodies, overexpression of IFN-α and/orIFN-β inducible genes, elevated levels of IP-10, elevated levels ofsCD40L, reduced levels of C3-derived C3b, reduced peripheral iNKT cellfrequencies, defective B cell-mediated stimulation of iNKT cells,altered CD1d expression on B cells, reduced numbers of naturalregulatory T cells (Treg), altered level of C-reactive protein,overexpression of mRNA for IL-4, overexpression of mRNA for IL-21, andelevated serium anti-collagen level. In some embodiments, the symptom isoverexpression of IFN-α, TNF-α, IL-6, IL-8, or IL-1. In one embodiment,the symptom is overexpression of IFN-α. In one embodiment, the symptomis overexpression of IL-6. In some embodiments, the symptom isoverexpression of mRNA for IL-4 or overexpression of mRNA for IL-21. Insome embodiments, the symptom is elevated serium anti-collagen level.

As used herein, and unless otherwise specified, to “decrease,”“ameliorate,” “reduce,” “inhibit,” “treat” (or the like) asthma or asymptom associated with asthma includes reducing the severity and/orfrequency of one or more symptoms of asthma, as well as preventingasthma and/or one or more symptoms of asthma (e.g., by reducing theseverity and/or frequency of flares of symptoms).

In some embodiments, the symptom is reduced by at least about 2%, atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 40%, atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, or at least about 95% relative to a controllevel. The control level includes any appropriate control as known inthe art. For example, the control level can be the pre-treatment levelin the sample or subject treated, or it can be the level in a controlpopulation (e.g., the level in subjects who do not have asthma or thelevel in samples derived from subjects who do not have asthma). In someembodiments, the decrease is statistically significant, for example, asassessed using an appropriate parametric or non-parametric statisticalcomparison.

In certain embodiments, the subject is an animal model of asthma, ahuman with asthma, or a subject (e.g., a human) at risk for developingasthma. In some embodiments, the subject is a human who has a familyhistory of asthma, who carries a gene associated with asthma, who ispositive for a biomarker associated with asthma, or a combinationthereof. In some embodiments, the subject has been diagnosed withasthma. In some embodiments, the subject has one or more signs orsymptoms associated with asthma. In some embodiments, the subject is atrisk for developing asthma (e.g., the subject carries a gene that,individually, or in combination with other genes or environmentalfactors, is associated with development of asthma).

In one embodiment, the subject has been previously diagnosed of asthmaor has episodic symptoms of airflow obstruction (e.g., wheezing and/orchest tightness) for at least 1 week, 2 weeks, 1 month, 2 months, 3months, 6 months, 9 months, 12 months before a compound provided herein(e.g., Compound 1), or an enantiomer, a mixture of enantiomers, or amixture of two or more diastereomers thereof, or a pharmaceuticallyacceptable form thereof, is administered. In one embodiment, the subjecthas been previously diagnosed of asthma or has episodic symptoms ofairflow obstruction (e.g., wheezing and/or chest tightness) for at least6 months before a compound provided herein (e.g., Compound 1), or anenantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof, isadministered.

In one embodiment, the subject has a forced expiratory volume in onesecond (FEV₁) value of at least 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%,55%, or 50% of a control value. In one embodiment, the subject has aforced expiratory volume in one second (FEV₁) value of at least 70% of acontrol value. In one embodiment, the control value may be calculatedbased on American Thoracic Society (ATS)/European Respiratory Society(ERS) standards.

In one embodiment, the subject has a positive response to a skin pricktest to an allergen. In one embodiment, the positive response means thatthe induration of skin test wheal is larger in diameter (e.g., at least2 mm larger) than the diameter of the control wheal. The allergen can beany allergen provided herein or known in the art that can be used in thediagnosis or determining status of asthma.

In one embodiment, the subject has an early-phase asthmatic response(EAR) of at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% toan inhaled allergen challenge. In one embodiment, the subject has anearly-phase asthmatic response of at least 20% to an inhaled allergenchallenge. In one embodiment, the EAR response is a decrease frompre-challenge in FEV₁ on 2 consecutive occasions within 0 to <3 hours oflast allergen challenge.

In one embodiment, the subject has a late-phase asthmatic response (LAR)of at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% to aninhaled allergen challenge. In one embodiment, the subject has alate-phase asthmatic response of at least 15% to an inhaled allergenchallenge. In one embodiment, the LAR response is a decrease frompre-challenge in FEV₁ on 2 consecutive occasions within 3 to 10 hours oflast allergen challenge.

In one embodiment, the subject has an early-phase asthmatic response ofat least 20% and a late-phase asthmatic response of at least 15% to aninhaled allergen challenge. The inhaled allergen can be any inhaledallergen provided herein or known in the art that can be used in thediagnosis or determining status of asthma.

In one embodiment, the subject exhibits an elevated level of C-reactiveprotein. In one embodiment, the subject exhibits an elevated level ofC-reactive protein of at least 1.0 mg/L. In one embodiment, the subjectexhibits an elevated level of C-reactive protein of at least 7 mg/L.

In some embodiments, the subject exhibits elevated levels of antinuclearantibodies (e.g., anti-Smith antibodies, anti-double stranded DNA(dsDNA) antibodies, anti-U1 RNP, SS-a (or anti-Ro), SS-b (or anti-La)),antiphospholipid antibodies, anti-ss DNA antibodies, anti-histoneantibodies, or anticardiolipin antibodies. In some embodiments, thesubject exhibits elevated levels of anti-dsDNA antibodies. In someembodiments, the subject exhibits elevated levels of anti-Sm antibodies.

In some embodiments, the subject exhibits autoantibodies against one ormore antigens that are known to be associated with asthma or with asthmasubtypes. In some embodiments, the subject exhibits autoantibodiesagainst Sm/anti-RNP or Ro/La autoantigens.

The levels of antibodies associated with asthma can be assessed usingany suitable method, e.g., methods known in the art, e.g., indirectimmunofluorescence. In some embodiments, the methods disclosed hereinreduce or prevent an increase in the levels of one or more of theforegoing antibodies.

In some embodiments, the subject exhibits elevated levels of IFN-α,TNF-α, IL-6, IL-8, or IL-1. In one embodiment, the subject exhibits anelevated level of IFN-α. In another embodiment, the subject exhibits anelevated level of IL-6. In another embodiment, the subject exhibits anelevated level of mRNA for IL-4 or IL-21.

In some embodiments, the subject has a mutation (e.g., an SNP) in a geneassociated with asthma. In one embodiment, the gene is selected fromSTAT4, IRF5, BANK1, ITGAM, PD1, FAM167A-BLK, IRF5-TNP03, KIAA1542,TNFA1P3, XKR6, 1q25.1, PXK, ATG5, ICA1, XKR6, LYN and SCUB2 or acombination thereof. In some embodiments, the subject carries the DR3and DQ2 variants, or the DR2 and DQ6 variants of HLA class II genes. Insome embodiments, the subject has a deficiency in one or more complementproteins, e.g. a deficiency of a complement protein coded by the C4A orC2 genes on chromosome 6, or the C1r and C1s genes on chromosome 12.

In some embodiments, the subject exhibits excessive PI3K activity orabnormal activity (e.g., excessive or reduced activity) of one or morecomponents of the PI3K signaling pathway (e.g., Akt (PKB), mTOR, a Teckinase (e.g., Btk, Itk, Tec), phospholipase C, PDK1, PKCs, NFκB, Rac GEF(e.g., Vav-1), or Rac).

In some embodiments, the subject is an animal model of asthma providedherein or known in the art. Examples include, but are not limited to,the murine lipopolysaccharide (LPS) induced pulmonary inflammationmodel, and the murine ovalbumin-induced allergic airway inflammationmodel.

In some embodiments, the subject has been previously treated for asthma.In some embodiments, the subject has been previously treated for asthmabut is non-responsive to standard therapies. Thus, in one embodiment,provided herein is a method of treating, preventing, and/or managingasthma in a subject, comprising administering an effective amount of acompound provided herein (e.g., Compound 1), or an enantiomer, a mixtureof enantiomers, or a mixture of two or more diastereomers thereof, or apharmaceutically acceptable form thereof, to a subject in need thereof,wherein the subject has been previously administered a therapy forasthma.

In some embodiments, the subject has not been previously treated forasthma.

In one embodiment, without being limited by any particular theory,administering an effective amount of a compound provided herein (e.g.,Compound 1), or an enantiomer, a mixture of enantiomers, or a mixture oftwo or more diastereomers thereof, or a pharmaceutically acceptable formthereof, does not result in, or results in reduced, one or more commonside effects of asthma treatment. The common side effects of asthmatreatment include, but are not limited to, oral candidiasis, thrush,dysphonia (hoarseness), reflex cough, bronchospasm, poor growth,decreased bone density, disseminated varicella infection (chickenpoxthat spreads to organs), easy bruising, cataracts, glaucoma, adrenalgland suppression, stomach upset, headache, liver test abnormalities,skin rashes, Churg Strauss syndrome, bad taste in month, cough, itching,sore throat, sneezing, stuffy nose, shortness of breath, wheezing, viralillness, upper respiratory tract infections, sinusitis, feeling dizzy orfaint, hives, changes in voice, swelling of the tongue, or difficulty inswallowing.

In some embodiments, the side effect is reduced by at least about 2%, atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 40%, atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, or at least about 95% relative to a controllevel. The control level includes any appropriate control as known inthe art. For example, the control level can be the side effect level inthe subject treated with other asthma therapies (e.g., Xolair, CromolynSodium, Nedocromil, Montelukast, and prednisone). In some embodiments,the decrease is statistically significant, for example, as assessedusing an appropriate parametric or non-parametric statisticalcomparison.

In one embodiment, the regression of asthma is a decrease (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% decrease) in thelevel of maximal decrease from pre-allergen challenge in FEV₁ followingallergen challenge. The level of maximal decrease from pre-allergenchallenge in FEV₁ following allergen challenge can be measured in EAR orLAR.

In one embodiment, the regression of asthma is a decrease (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% decrease) in areaunder the curve (AUC) of FEV₁ following allergen challenge.

In one embodiment, the regression of asthma is an increase (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% increase) in theamount of methacholine that is required to induce a 20% fall in FEV_(I)(PC₂₀) following allergen challenge.

In one embodiment, the regression of asthma is a decrease (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% decrease) inexhaled nitric oxide level of the subject.

In one embodiment, the regression of asthma is a decrease (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% decrease) in theC-reactive protein (CRP) level of the subject.

In one embodiment, the regression of asthma is a decrease (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% decrease) in whiteblood cell count and/or differential cell count in induced sputum of thesubject after allergen challenge.

5.6. Combination Therapy

In some embodiments, the compound provided herein is administered incombination with one or more other therapies. In one embodiment,provided herein are methods for combination therapies in which an agentknown to modulate other pathways, or other components of the samepathway, or even overlapping sets of target enzymes are used incombination with a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof. In one aspect, such therapy includes, but is not limited to,the combination of the subject compound with chemotherapeutic agents,therapeutic antibodies, and/or radiation treatment, to provide asynergistic or additive therapeutic effect.

By “in combination with,” it is not intended to imply that the othertherapy and the PI3K modulator must be administered at the same timeand/or formulated for delivery together, although these methods ofdelivery are within the scope of this disclosure. The compound providedherein can be administered concurrently with, prior to (e.g., 5 minutes,15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours,12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 12 weeks, or 16 weeksbefore), or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6weeks, 8 weeks, 12 weeks, or 16 weeks after), one or more othertherapies (e.g., one or more other additional agents). In general, eachtherapeutic agent will be administered at a dose and/or on a timeschedule determined for that particular agent. The other therapeuticagent can be administered with the compound provided herein in a singlecomposition or separately in a different composition. Triple therapy isalso contemplated herein.

In general, it is expected that additional therapeutic agents employedin combination be utilized at levels that do not exceed the levels atwhich they are utilized individually. In some embodiments, the levelsutilized in combination will be lower than those utilized individually.

In some embodiments, the compound provided herein is a first linetreatment for cancer or hematologic malignancy, i.e., it is used in asubject who has not been previously administered another drug or therapyintended to treat cancer or hematologic malignancy or one or moresymptoms thereof.

In other embodiments, the compound provided herein is a second linetreatment for cancer or hematologic malignancy, i.e., it is used in asubject who has been previously administered another drug or therapyintended to treat cancer or hematologic malignancy or one or moresymptoms thereof.

In other embodiments, the compound provided herein is a third or fourthline treatment for cancer or hematologic malignancy, i.e., it is used ina subject who has been previously administered two or three other drugsor therapies intended to treat cancer or hematologic malignancy or oneor more symptoms thereof.

In embodiments where two agents are administered, the agents can beadministered in any order. For example, the two agents can beadministered concurrently (i.e., essentially at the same time, or withinthe same treatment) or sequentially (i.e., one immediately following theother, or alternatively, with a gap in between administration of thetwo). In some embodiments, the compound provided herein is administeredsequentially (i.e., after the first therapeutic).

In one aspect, a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein, can presentsynergistic or additive efficacy when administered in combination withagents that inhibit IgE production or activity. Such combination canreduce the undesired effect of high level of IgE associated with the useof one or more PI3K-δ inhibitors, if such effect occurs. This can beparticularly useful in treatment of autoimmune and inflammatorydisorders (AIID) such as rheumatoid arthritis. Additionally, theadministration of PI3K-δ, PI3K-γ, or PI3K-δ/γ inhibitors as providedherein in combination with inhibitors of mTOR can also exhibit synergythrough enhanced inhibition of the PI3K pathway.

In a separate but related aspect, provided herein is a combinationtreatment of a disease associated with PI3K-δ comprising administeringto a subject in need thereof a PI3K-δ inhibitor and an agent thatinhibits IgE production or activity. Other exemplary PI3K-δ inhibitorsare applicable for this combination and they are described in, e.g.,U.S. Pat. No. 6,800,620, incorporated herein by reference. Suchcombination treatment is particularly useful for treating autoimmune andinflammatory diseases (AIID) including, but not limited to rheumatoidarthritis.

Agents that inhibit IgE production are known in the art and theyinclude, but are not limited to, one or more of TEI-9874,2-(4-(6-cyclohexyloxy-2-naphtyloxy)phenylacetamide)benzoic acid,rapamycin, rapamycin analogs (i.e., rapalogs), TORC1 inhibitors, TORC2inhibitors, and any other compounds that inhibit mTORC1 and mTORC2.Agents that inhibit IgE activity include, for example, anti-IgEantibodies such as for example Omalizumab and TNX-901.

For treatment of autoimmune diseases, a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein, can be used in combination with commonly prescribed drugsincluding, but not limited to, Enbrel®, Remicade®, Humira®, Avonex®, andRebif®. For treatment of respiratory diseases, the subject compounds, orpharmaceutically acceptable forms thereof, or pharmaceuticalcompositions, can be administered in combination with commonlyprescribed drugs including, but not limited to, Xolair®, Advair®,Singulair®, and Spiriva®.

The compounds as provided herein, or pharmaceutically acceptable forms(e.g., pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, orpharmaceutical compositions as provided herein, can be formulated oradministered in conjunction with other agents that act to relieve thesymptoms of inflammatory conditions such as encephalomyelitis, asthma,and the other diseases described herein. These agents includenon-steroidal anti-inflammatory drugs (NSAIDs), e.g., acetylsalicylicacid; ibuprofen; naproxen; indomethacin; nabumetone; tolmetin; etc.Corticosteroids are used to reduce inflammation and suppress activity ofthe immune system. An exemplary drug of this type is Prednisone.Chloroquine (Aralen) or hydroxychloroquine (Plaquenil) can also be usedin some individuals with lupus. They can be prescribed for skin andjoint symptoms of lupus. Azathioprine (Imuran) and cyclophosphamide(Cytoxan) suppress inflammation and tend to suppress the immune system.Other agents, e.g., methotrexate and cyclosporin are used to control thesymptoms of lupus. Anticoagulants are employed to prevent blood fromclotting rapidly. They range from aspirin at very low dose whichprevents platelets from sticking, to heparin/coumadin. Other compoundsused in the treatment of lupus include belimumab (Benlysta®).

In another aspect, provided herein is a pharmaceutical composition forinhibiting abnormal cell growth in a subject which comprises an amountof a compound provided herein, or a pharmaceutically acceptable form(e.g., pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, in combinationwith an amount of an anti-cancer agent (e.g., a chemotherapeutic agentor a biotherapeutic agent). Many chemotherapeutics are presently knownin the art and can be used in combination with a compound providedherein.

In some embodiments, the chemotherapeutic is selected from mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, anti-hormones,angiogenesis inhibitors, and anti-androgens. Non-limiting examples arechemotherapeutic agents, cytotoxic agents, and non-peptide smallmolecules such as Gleevec® (imatinib mesylate), Velcade® (bortezomib),Casodex™ (bicalutamide), Iressa® (gefitinib), Tarceva® (erlotinib), andAdriamycin® (doxorubicin) as well as a host of chemotherapeutic agents.Non-limiting examples of chemotherapeutic agents include alkylatingagents such as thiotepa and cyclosphosphamide (CYTOXAN™); alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines suchas benzodopa, carboquone, meturedopa, and uredopa; ethylenimines andmethylamelamines including altretamine, triethylenemelamine,trietylenephosphoramide, triethylenethiophosphoramide andtrimethylolomelamine; BTK inhibitors such as ibrutinib (PCI-32765),AVL-292, Dasatinib, LFM-AI3, ONO-WG-307, and GDC-0834; HDAC inhibitorssuch as vorinostat, romidepsin, panobinostat, valproic acid, belinostat,mocetinostat, abrexinostat, entinostat, SB939, resminostat, givinostat,CUDC-101, AR-42, CHR-2845, CHR-3996, 4SC-202, CG200745, ACY-1215 andkevetrin; EZH2 inhibitors such as, but not limited to, EPZ-6438(N-(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methyl-4′-(morpholinomethyl)-[1,1′-biphenyl]-3-carboxamide),GSK-126((S)-1-(sec-butyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-methyl-6-(6-(piperazin-1-yl)pyridin-3-yl)-1H-indole-4-carboxamide),GSK-343(1-Isopropyl-N-(6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl)-6-(2-(4-methylpiperazin-1-yl)pyridine-4-yl)-1H-indazole-4-carboxamide),E11, 3-deazaneplanocin A (DNNep,5R-(4-amino-1H-imidazo[4,5-c]pyridin-1-yl)-3-(hydroxymethyl)-3-cyclopentene-1S,2R-diol),small interfering RNA (siRNA) duplexes targeted against EZH2 (S. M.Elbashir et al., Nature 411:494-498 (2001)), isoliquiritigenin, andthose provided in, for example, U.S. Publication Nos. 2009/0012031,2009/0203010, 2010/0222420, 2011/0251216, 2011/0286990, 2012/0014962,2012/0071418, 2013/0040906, and 2013/0195843, all of which areincorporated herein by reference; JAK/STAT inhibitors such aslestaurtinib, tofacitinib, ruxolitinib, pacritinib, CYT387, baricitinib,GLPG0636, TG101348, INCB16562, CP-690550, and AZD1480; PKC-β inhibitorsuch as Enzastaurin; SYK inhibitors such as, but not limited to,GS-9973, PRT 062607, R406,(S)-2-(2-((3,5-dimethylphenyl)amino)pyrimidin-4-yl)-N-(1-hydroxypropan-2-yl)-4-methylthiazole-5-carboxamide,R112, GSK143, BAY61-3606, PP2, PRT 060318, R348, and those provided in,for example, U.S. Publication Nos. 2003/0113828, 2003/0158195,2003/0229090, 2005/0075306, 2005/0232969, 2005/0267059, 2006/0205731,2006/0247262, 2007/0219152, 2007/0219195, 2008/0114024, 2009/0171089,2009/0306214, 2010/0048567, 2010/0152159, 2010/0152182, 2010/0316649,2011/0053897, 2011/0112098, 2011/0245205, 2011/0275655, 2012/0027834,2012/0093913, 2012/0101275, 2012/0130073, 2012/0142671, 2012/0184526,2012/0220582, 2012/0277192, 2012/0309735, 2013/0040984, 2013/0090309,2013/0116260, and 2013/0165431, all of which are incorporated herein byreference; SYK inhibitor such as R788 (fostamatinib); SYK/JAK dualinhibitor such as PRT2070; nitrogen mustards such as bendamustine,chlorambucil, chlornaphazine, cholophosphamide, estramustine,ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride,melphalan, novembichin, phenesterine, prednimustine, trofosfamide,uracil mustard; nitrosureas such as carmustine, chlorozotocin,fotemustine, lomustine, nimustine, ranimustine; antibiotics such asaclacinomycins, actinomycin, authramycin, azaserine, bleomycins,cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin,chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycin C, mycophenolic acid, nogalamycin,olivomycins, peplomycin, porfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pralatrexate, pteropterin, trimetrexate; purine analogssuch as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine,carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine,floxuridine, androgens such as calusterone, dromostanolone propionate,epitiostanol, mepitiostane, testolactone; anti-adrenals such asaminoglutethimide, mitotane, trilostane; folic acid replenisher such asfolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinicacid; amsacrine; bestrabucil; bisantrene; edatrexate; defofamine;demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone;mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK.R™; razoxane;sizofiran; spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethyla-mine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (Ara-C); cyclophosphamide; thiotepa; taxanes, e.g.,paclitaxel (e.g., TAXOL™) and docetaxel (e.g., TAXOTERE™) and ABRAXANE®(paclitaxel protein-bound particles); retinoic acid; esperamicins;capecitabine; and pharmaceutically acceptable forms (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) of any of the above.Also included as suitable chemotherapeutic cell conditioners areanti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens including for example tamoxifen(Nolvadex™), raloxifene, aromatase inhibiting 4(5)-imidazoles,4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, andtoremifene (Fareston); and anti-androgens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine;6-thioguanine; mercaptopurine; methotrexate; platinum analogs such ascisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16);ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine;navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda;ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000;difluoromethylornithine (DMFO). Where desired, the compounds orpharmaceutical composition as provided herein can be used in combinationwith commonly prescribed anti-cancer drugs such as Herceptin®, Avastin®,Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE,abagovomab, acridine carboxamide, adecatumumab,17-N-allylamino-17-demethoxygeldanamycin, alpharadin, alvocidib,3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide,anthracenedione, anti-CD22 immunotoxins, antineoplastic, antitumorigenicherbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine,BIBW 2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine,CBV (chemotherapy), calyculin, crizotinib, cell-cycle nonspecificantineoplastic agents, dichloroacetic acid, discodermolide,elsamitrucin, enocitabine, epothilone, eribulin, everolimus, exatecan,exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen,IT-101, imexon, imiquimod, indolocarbazole, irofulven, laniquidar,larotaxel, lenalidomide, lucanthone, lurtotecan, mafosfamide,mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1,pawpaw, pixantrone, proteasome inhibitor, rebeccamycin, resiquimod,rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V,swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel,triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine,uramustine, vadimezan, vinflunine, ZD6126, and zosuquidar.

In some embodiments, the chemotherapeutic is selected from hedgehoginhibitors including, but not limited to IPI-926 (See U.S. Pat. No.7,812,164). Other suitable hedgehog inhibitors include, for example,those described and disclosed in U.S. Pat. No. 7,230,004, U.S. PatentApplication Publication No. 2008/0293754, U.S. Patent ApplicationPublication No. 2008/0287420, and U.S. Patent Application PublicationNo. 2008/0293755, the entire disclosures of which are incorporated byreference herein. Examples of other suitable hedgehog inhibitors includethose described in U.S. Patent Application Publication Nos. US2002/0006931, US 2007/0021493 and US 2007/0060546, and InternationalApplication Publication Nos. WO 2001/19800, WO 2001/26644, WO2001/27135, WO 2001/49279, WO 2001/74344, WO 2003/011219, WO2003/088970, WO 2004/020599, WO 2005/013800, WO 2005/033288, WO2005/032343, WO 2005/042700, WO 2006/028958, WO 2006/050351, WO2006/078283, WO 2007/054623, WO 2007/059157, WO 2007/120827, WO2007/131201, WO 2008/070357, WO 2008/110611, WO 2008/112913, and WO2008/131354, each incorporated herein by reference. Additional examplesof hedgehog inhibitors include, but are not limited to, GDC-0449 (alsoknown as RG3616 or vismodegib) described in, e.g., Von Hoff D. et al., NEngl. J. Med. 2009; 361(12):1164-72; Robarge K. D. et al., Bioorg MedChem Lett. 2009; 19(19):5576-81; Yauch, R. L. et al. (2009) Science 326:572-574; Sciencexpress: 1-3 (10.1126/science.1179386); Rudin, C. et al.(2009) New England J of Medicine 361-366 (10.1056/nejma0902903);BMS-833923 (also known as XL139) described in, e.g., in Siu L. et al.,J. Clin. Oncol. 2010; 28:15s (suppl; abstr 2501); and National Instituteof Health Clinical Trial Identifier No. NCT006701891; LDE-225 described,e.g., in Pan S. et al., ACS Med. Chem. Lett., 2010; 1(3): 130-134;LEQ-506 described, e.g., in National Institute of Health Clinical TrialIdentifier No. NCT01106508; PF-04449913 described, e.g., in NationalInstitute of Health Clinical Trial Identifier No. NCT00953758; Hedgehogpathway antagonists disclosed in U.S. Patent Application Publication No.2010/0286114; SMOi2-17 described, e.g., U.S. Patent ApplicationPublication No. 2010/0093625; SANT-1 and SANT-2 described, e.g., inRominger C. M. et al., J. Pharmacol. Exp. Ther. 2009; 329(3):995-1005;1-piperazinyl-4-arylphthalazines or analogues thereof, described inLucas B. S. et al., Bioorg. Med. Chem. Lett. 2010; 20(12):3618-22.

Other hormonal therapy and chemotherapeutic agents include, but are notlimited to, anti-estrogens (e.g. tamoxifen, raloxifene, and megestrolacetate), LHRH agonists (e.g. goserelin and leuprolide), anti-androgens(e.g. flutamide and bicalutamide), photodynamic therapies (e.g.vertoporfin (BPD-MA), phthalocyanine, photosensitizer Pc4, anddemethoxy-hypocrellin A (2BA-2-DMHA)), nitrogen mustards (e.g.cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, estramustine,and melphalan), nitrosoureas (e.g. carmustine (BCNU) and lomustine(CCNU)), alkylsulphonates (e.g. busulfan and treosulfan), triazenes(e.g. dacarbazine, temozolomide), platinum containing compounds (e.g.cisplatin, carboplatin, oxaliplatin), vinca alkaloids (e.g. vincristine,vinblastine, vindesine, and vinorelbine), taxoids or taxanes (e.g.paclitaxel or a paclitaxel equivalent such as nanoparticle albumin-boundpaclitaxel (Abraxane), docosahexaenoic acid bound-paclitaxel(DHA-paclitaxel, Taxoprexin), polyglutamate bound-paclitaxel(PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX), thetumor-activated prodrug (TAP) ANG1005 (Angiopep-2 bound to threemolecules of paclitaxel), paclitaxel-EC-1 (paclitaxel bound to theerbB2-recognizing peptide EC-1), and glucose-conjugated paclitaxel,e.g., 2′-paclitaxel methyl 2-glucopyranosyl succinate; docetaxel,taxol), epipodophyllins (e.g. etoposide, etoposide phosphate,teniposide, topotecan, 9-aminocamptothecin, camptoirinotecan,irinotecan, crisnatol, mytomycin C), anti-metabolites, DHFR inhibitors(e.g. methotrexate, dichloromethotrexate, trimetrexate, edatrexate), IMPdehydrogenase inhibitors (e.g. mycophenolic acid, tiazofurin, ribavirin,and EICAR), ribonuclotide reductase inhibitors (e.g. hydroxyurea anddeferoxamine), uracil analogs (e.g. 5-fluorouracil (5-FU), floxuridine,doxifluridine, raltitrexed, tegafur-uracil, capecitabine), cytosineanalogs (e.g. cytarabine (ara C, cytosine arabinoside), andfludarabine), purine analogs (e.g. mercaptopurine and thioguanine),Vitamin D3 analogs (e.g. EB 1089, CB 1093, and KH 1060), isoprenylationinhibitors (e.g. lovastatin), dopaminergic neurotoxins (e.g.1-methyl-4-phenylpyridinium ion), cell cycle inhibitors (e.g.staurosporine), actinomycin (e.g. actinomycin D, dactinomycin),bleomycin (e.g. bleomycin A2, bleomycin B2, peplomycin), anthracyclines(e.g. daunorubicin, doxorubicin, pegylated liposomal doxorubicin,idarubicin, epirubicin, pirarubicin, zorubicin, mitoxantrone), MDRinhibitors (e.g. verapamil), Ca2+ ATPase inhibitors (e.g. thapsigargin),thalidomide, lenalidomide (REVLIMID®), tyrosine kinase inhibitors (e.g.,axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTIN™,AZD2171), dasatinib (SPRYCEL®, BMS-354825), erlotinib (TARCEVA®),gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib(TYKERB®, TYVERB®), lestaurtinib (CEP-701), neratinib (HKI-272),nilotinib (TASIGNA®), semaxanib (semaxinib, SU5416), sunitinib (SUTENT®,SU11248), toceranib (PALLADIA®), vandetanib (ZACTIMA®, ZD6474),vatalanib (PTK787, PTK/ZK), trastuzumab (HERCEPTIN®), bevacizumab(AVASTIN®), rituximab (RITUXAN®), cetuximab (ERBITUX®), panitumumab(VECTIBIX®), ranibizumab (Lucentis®), sorafenib (NEXAVAR®), everolimus(AFINITOR®), alemtuzumab (CAMPATH®), gemtuzumab ozogamicin (MYLOTARG®),temsirolimus (TORISEL®), ENMD-2076, PCI-32765, AC220, dovitinib lactate(TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903,PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120(VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154,CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, and/orXL228), proteasome inhibitors (e.g., bortezomib (Velcade)), mTORinhibitors (e.g., rapamycin, temsirolimus (CCI-779), everolimus(RAD-001), ridaforolimus, AP23573 (Ariad), AZD8055 (AstraZeneca), BEZ235(Novartis), BGT226 (Norvartis), XL765 (Sanofi Aventis), PF-4691502(Pfizer), GDC0980 (Genetech), SF1126 (Semafoe) and OSI-027 (OSI)),oblimersen, gemcitabine, carminomycin, leucovorin, pemetrexed,cyclophosphamide, dacarbazine, procarbazine, prednisolone,dexamethasone, camptothecin, plicamycin, asparaginase, aminopterin,methopterin, porfiromycin, melphalan, leurosidine, leurosine,chlorambucil, trabectedin, procarbazine, discodermolide, carminomycin,aminopterin, and hexamethyl melamine.

Exemplary biotherapeutic agents include, but are not limited to,interferons, cytokines (e.g., tumor necrosis factor, interferon α,interferon γ), vaccines, hematopoietic growth factors, monoclonalserotherapy, immuno-stimulants and/or immuno-modulatory agents (e.g.,IL-1, 2, 4, 6, 7, 12, 15, or 21), immune cell growth factors (e.g.,GM-CSF) and antibodies (e.g. Herceptin (trastuzumab), T-DM1, AVASTIN(bevacizumab), ERBITUX (cetuximab), Vectibix (panitumumab), Rituxan(rituximab), Bexxar (tositumomab), or Perjeta (pertuzumab)).

In some embodiments, the biotherapeutic agent is an immunotherapeuticagent, e.g., a cancer vaccine e.g., a tumor vaccine. Exemplary cancervaccines include Aduro (GVAX); Advaxis (ADXS11-001, ADXS31-001,ADXS31-164, ADXS31-142 (ADXS-PSA)); ALVAC-CEA vaccine; Avax Technologies(AC Vaccine); Amgen (talimogene laherparepvec); Biovest International(BiovaxID in phase III); Bavarian Nordic (PROSTVAC); CelldexTherapeutics (CDX110, CDX1307 and CDX1401); The Center of MolecularImmunology (CimaVax-EGF); CureVac develops mRNA-based cancerimmunotherapies; CV9104; Dendreon Corp (Neuvenge); Galena Biopharma(NeuVax); Antigen Express (Ae-37); Geron Corporation (GRNVAC1);GlobeImmune (Tarmogens, GI-4000, GI-6207, GI-6301); Heat Biologics(ImPACT Therapy); Immatics biotechnologies (IMA901); Merck (Stimuvax);Panacela Labs, Inc. (MOBILAN Adenovirus-based treatment); Prima BioMed(Cvac); Scancell Holdings (SCIB 1).

In embodiments, the biotherapeutic agent is a cellular therapy, e.g.,dendritic cell therapy or a chimeric T cell therapy such as CART.Dendritic cell therapy can comprise loading dendritic cells with anantigen obtained from a patient's tumor, then administering thedendritic cells to the patient in order to sensitize the patient's own Tcells to the tumor antigens. Chimeric antigen receptors (CARs) areengineered receptors that can be used to confer tumor specificity to a Tcell. CARS have been generated with specificity for α-folate receptor,CAIX, CD19, CD20, CD22, CD30, CD33, CD44v7/8, CEA, EGP-2, EGP-40,erb-B2, erb-B 2,3,4, FBP, Fetal acethylcholine receptor, FD2, Her2/neu,IL13R-a2, KDR, k-light chain, LeY, L1 cell adhesion molecule, MAGE-A1,mesothelin, CMV-infected cells, MUC1, NKG2D ligands, oncofetal antigenh5T4, PSCA, PSMA, TAA, TAG-72, and VEGF-R2.

In one embodiment, the biotherapeutic agent is an anti-CD37 antibodysuch as, but not limited to, IMGN529, K7153A and TRU-016. In anotherembodiment, the biotherapeutic agent is an anti-CD20 antibody such as,but not limited to, ¹³¹I tositumomab, ⁹⁰Y ibritumomab, ¹¹¹I ibritumomab,obinutuzumab (GAZYVA), and ofatumumab. In another embodiment, thebiotherapeutic agent is an anti-CD52 antibody such as, but not limitedto, alemtuzumab.

In some embodiments, the chemotherapeutic is selected from HSP90inhibitors. The HSP90 inhibitor can be a geldanamycin derivative, e.g.,a benzoquinone or hygroquinone ansamycin HSP90 inhibitor (e.g., IPI-493and/or IPI-504). Non-limiting examples of HSP90 inhibitors includeIPI-493, IPI-504, 17-AAG (also known as tanespimycin or CNF-1010),BIIB-021 (CNF-2024), BIIB-028, AUY-922 (also known as VER-49009),SNX-5422, STA-9090, AT-13387, XL-888, MPC-3100, CU-0305, 17-DMAG,CNF-1010, Macbecin (e.g., Macbecin I, Macbecin II), CCT-018159,CCT-129397, PU-H71, or PF-04928473 (SNX-2112).

In some embodiments, the chemotherapeutic is selected from PI3Kinhibitors (e.g., including those PI3K inhibitors provided herein andthose PI3K inhibitors not provided herein). In some embodiment, the PI3Kinhibitor is an inhibitor of delta and gamma isoforms of PI3K. In someembodiment, the PI3K inhibitor is an inhibitor of delta isoform of PI3K.In some embodiment, the PI3K inhibitor is an inhibitor of gamma isoformof PI3K. In some embodiments, the PI3K inhibitor is an inhibitor ofalpha isoform of PI3K. In other embodiments, the PI3K inhibitor is aninhibitor of one or more alpha, beta, delta and gamma isoforms of PI3K.Exemplary PI3K inhibitors that can be used in combination are describedin, e.g., WO 09/088990, WO 09/088086, WO 2011/008302, WO 2010/036380, WO2010/006086, WO 09/114870, WO 05/113556; US 2009/0312310, and US2011/0046165, each incorporated herein by reference. Additional PI3Kinhibitors that can be used in combination with the pharmaceuticalcompositions, include but are not limited to, RP-6530, TG 100-115,RV1729, AMG-319, GSK 2126458, GDC-0980, GDC-0941, Sanofi XL147, XL499,XL756, XL147, PF-4691502, BKM 120, CAL-101 (GS-1101), CAL 263, SF1126,PX-886, and a dual PI3K inhibitor (e.g., Novartis BEZ235). In oneembodiment, the PI3K inhibitor is an isoquinolinone. In one embodiment,the PI3K inhibitor is RP-6530, which has the chemical name:(S)-2-(1-((9H-purin-6-yl)amino)propyl)-3-(3-fluorophenyl)-4H-chromen-4-one.In one embodiment, the PI3K inhibitor is TG 100-115, which has thechemical name: 6,7-Bis(3-hydroxyphenyl)pteridine-2,4-diamine. In oneembodiment, the PI3K inhibitor is RV1729, which has the chemical name:6-(2-((4-amino-3-(3-hydroxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-3-(2-chlorobenzyl)-4-oxo-3,4-dihydroquinazolin-5-yl)-N,N-bis(2-methoxyethyl)hex-5-ynamide.

Without being bound by any particular theory, it is believed that therole of each PI3K isoform is critically dependent on cell type andupstream initiating signals, and therefore pharmacologic inhibition ofspecific isoforms can lead to different physiologic outcomes. PI3K is alipid kinase existing in multiple isoforms that have central roles inthe regulation of important cellular processes, including cell growthand survival. Puri et al., Frontiers in Immunology. 2012, 3: 256. PI3K-δand PI3K-γ are both expressed in CLL and NHL tumor cells. Signalingthrough PI3K is critical for supporting the growth and survival of thesemalignancies as they mediate intracellular BCR signaling and promoteinteractions between the tumor cells and their microenvironment. Puri etal., Frontiers in Immunology. 2012, 3: 256.

The specific functions of PI3K-δ in malignant B cells support therationale for it to be a therapeutic target to control these diseases.PI3K-δ inhibition disrupts malignant cell interaction with the stromalmicroenvironment, thereby short-circuiting chemokine-mediatedstimulation of CLL and other B-cell malignancies, priming cells forapoptosis by pharmacologic or natural stimuli. Pharmacologic inhibitionof PI3K-δ reduces disease activity in various models of B-cell-derivedmalignancies, including CLL and B-cell lymphomas. PI3K-δ inhibitionimproves the therapeutic potential of other antitumor agents in variouspreclinical models of B-cell malignancy, including CLL. Lannutti et al.,Blood. 2011, 117, 591-594.

The role of PI3K-γ in cells that maintain the malignant B-cellmicroenvironment creates potential for therapeutic inhibition of PI3K-γto control these diseases. PI3K-γ plays a role in T-cell activation andmigration and GPCR-associated chemokine signaling. Reif et al. JImmunol. 2004; 173:2236-2240. PI3K-γ also mediates adhesion andtrafficking of tumor-associated macrophages (TAMs). Reif et al. JImmunol. 2004; 173:2236-2240. Hasan et al., Int Immunopharmacology.2010, 10, 1017-1021; Laffargue et al., Immunity. 2002, 16, 441-451.There is dynamic interplay or “cross-talk” between PI3K-δ and PI3K-γ inessential cellular activities in malignant cells. In certain tumortypes, PI3K-γ can promote tumorigenesis in the absence of PI3K-δ.Subramaniam et al., Cancer Cell. 2012, 21, 459-472.

In certain tumor types, dual isoform inhibition may be necessary foroptimal tumor growth inhibition in preclinical models. Subramaniam etal., Cancer Cell. 2012, 21, 459-472. As shown in the examples, in somecell lines (e.g., NHL (e.g., follicular lymphoma), DLBCL, mantle cell,multiple myeloma, T-cell lymphoma), combined inhibition of PI3K-δ andPI3K-γ shows greater growth inhibition than inhibition of either isoformalone.

In one embodiment, the PI3K-delta selective compound is GSK-2269557(2-(6-(1H-indol-4-yl)-1H-indazol-4-yl)-5-((4-isopropylpiperazin-1-yl)methyl)oxazole),GS9820 (CAL-120,(S)-2-(1-((9H-purin-6-yl)amino)ethyl)-6-fluoro-3-phenylquinazolin-4(3H)-one),GS-1101(5-fluoro-3-phenyl-2-([S)]-1-[9H-purin-6-ylamino]-propyl)-3H-quinazolin-4-one),AMG319, or TGR-1202((S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one),or a mixture thereof. In one embodiment, the PI3K-delta selectivecompound is GS1101.

In one embodiment, the PI3K-delta selective compound is a PI3K-deltainhibitor as described in WO 2005/113556, the entirety of which isincorporated herein by reference. In one embodiment, the PI3K-deltaselective compound is Compound No. 113 or 107 as described in2005/113556.

In one embodiment, the PI3K-delta selective compound is a PI3K-deltacompound as described in WO2014/006572, the entirety of which isincorporated herein by reference. In one embodiment, the PI3K-deltaselective compound is a PI3K-delta inhibitor has an alpha/deltaselectivity ratio of greater than about 100, greater than about 250,greater than about 500, greater than about 750, or greater than about1000. In one embodiment, the PI3K-delta selective compound is aPI3K-delta inhibitor has a beta/delta selectivity ratio of greater thanabout 10, greater than about 20, greater than about 30, greater thanabout 40, or greater than about 50. In one embodiment, the PI3K-deltaselective compound is a PI3K-delta inhibitor has a gamma/deltaselectivity ratio of greater than about 1, greater than about 10,greater than about 25, greater than about 30, or greater than about 50.In one embodiment, the PI3K-delta selective compound is Compound Nos.A1, A2, B, B1 or B2 as described in WO2014/006572. In one embodiment,the PI3K-delta selective compound is Compound No. B1 as described inWO2014/006572.

In one embodiment, the PI3K-delta selective compound is a PI3K-deltacompound as described in WO 2013/032591, the entirety of which isincorporated herein by reference. In one embodiment, the PI3K-deltaselective compound is a compound of Formula (I) as described in WO2013/032591. In one embodiment, the PI3K-delta selective compound is acompound described in WO 2013/032591 with a IC₅₀ (nM) for the PI3K-deltaisoform of less than 100 nM and a IC₅₀ (nM) for the PI3K-alpha, beta, orgamma of greater than about 100 nM, greater than about 1 μM, or greaterthan about 10 μM. In one embodiment, the PI3K-delta selective compoundhas an alpha/delta selectivity ratio, a beta/delta selectivity ratio, ora gamma/delta selectivity ratio of greater than 1, greater than about10, or greater than about 100. In one embodiment, the PI3K-deltaselective compound is Compound Nos. 13, 30, 41, 55, 57, 124, 167, 183,185, 187, 191, 196, 226, 230, 232, 234, 235, 326, 327, 328, 333, 334,336, 337, 338, 356, 359, 378, 439, 440, 443, or 455, as described in WO2013/032591. In one embodiment, the PI3K-delta selective compound isCompound Nos. 183, 230, 234, 235, 326, 333, 336, 337, 338, or 359, asdescribed in WO 2013/032591. In one embodiment, the PI3K-delta selectivecompound is Compound No. 359 as described in WO 2013/032591.

In one embodiment, provided herein are pharmaceutical compositionscomprising a compound of Formula (I′), (A′), (I), or (A), or apharmaceutically acceptable form thereof, and a PI3K-delta selectivecompound, wherein the PI3K-delta selective compound is GSK-2269557,GS-9820, GS-1101 (Cal-101 or idelalisib), AMG319, or TGR-1202, or amixture thereof. In one embodiment, the PI3K-delta selective compound isGS1101. In one embodiment, the composition is synergistic in treating orpreventing a PI3K mediated disorder. In one embodiment, the PI3K-deltaselective compound is a compound described in WO2011/146882, theentirety of which is incorporated herein by reference. In oneembodiment, the PI3K-delta selective compound is a compound described inWO2011/146882 with a IC₅₀ (nM) for the PI3K-delta isoform of less than100 nM and a IC₅₀ (nM) for the PI3K-alpha, beta, or gamma of greaterthan about 100 nM, greater than about 1 μM, or greater than about 10 μM.In one embodiment, the PI3K-delta selective compound has an alpha/deltaselectivity ratio, a beta/delta selectivity ratio, or a gamma/deltaselectivity ratio of greater than 1, greater than about 10, or greaterthan about 100. In one embodiment, the PI3K-delta selective compound isCompound No. 69 as described in WO2011/146882.

In one embodiment, the PI3K-delta selective compound is a compounddescribed in WO2013/012915, the entirety of which is incorporated hereinby reference. In one embodiment, the PI3K-delta selective compound is acompound described in WO2013/012915 with a IC₅₀ (nM) for the PI3K-deltaisoform of less than 100 nM and a IC₅₀ (nM) for the PI3K-alpha, beta, orgamma of greater than about 100 nM, greater than about 1 μM, or greaterthan about 10 μM. In one embodiment, the PI3K-delta selective compoundhas an alpha/delta selectivity ratio, a beta/delta selectivity ratio, ora gamma/delta selectivity ratio of greater than 1, greater than about10, or greater than about 100. In one embodiment, the PI3K-deltaselective compound is Compound No. I-41 or I-106 as described inWO2013/012915.

In one embodiment, the PI3K-delta selective compound is a compounddescribed in WO2013/012918, the entirety of which is incorporated hereinby reference. In one embodiment, the PI3K-delta selective compound is acompound described in WO2013/012918 with a IC₅₀ (nM) for the PI3K-deltaisoform of less than 100 nM and a IC₅₀ (nM) for the PI3K-alpha, beta, orgamma of greater than about 100 nM, greater than about 1 μM, or greaterthan about 10 μM. In one embodiment, the PI3K-delta selective compoundhas an alpha/delta selectivity ratio, a beta/delta selectivity ratio, ora gamma/delta selectivity ratio of greater than 1, greater than about10, or greater than about 100. In one embodiment, the PI3K-deltaselective compound is Compound No. 19, 28, 37, 38, 51, 59, 60, 89, 92,103, 106, 107, 108, or 109 as described in WO2013/012918. In oneembodiment, the PI3K-delta selective compound is Compound No. 103 or 106as described in WO2013/012918.

In one embodiment, provided herein are methods of treating or preventinga PI3K mediated disorder in a subject, comprising administering to thesubject a therapeutically effective amount of a compound of Formula(I′), (A′), (I), or (A), or a pharmaceutically acceptable form thereof,in combination with a PI3K-delta selective compound, wherein thePI3K-delta selective compound is GSK-2269557, GS-9820, GS-1101(Cal-101), AMG319, or TGR-1202, or a mixture thereof. In one embodiment,provided herein are methods of enhancing a PI3K-delta selective compoundtreatment of a PI3K mediated disorder in a subject comprisingadministering a compound of Formula (I′), (A′), (I), or (A), or apharmaceutically acceptable form thereof, in combination with the PI3Kselective delta compound, wherein the PI3K-delta selective compound isGSK-2269557, GS9-820, GS-1101 (Cal-101), AMG319, or TGR-1202, or amixture thereof. In one embodiment, the PI3K-delta selective compound isGS1101. In one embodiment, the administering a compound of Formula (I′),(A′), (I), or (A), or a pharmaceutically acceptable form thereof, incombination with the PI3K-delta selective compound provides synergisticeffect.

Also provided herein are methods of inhibiting growth of a cellcomprising contacting the cell with a compound of Formula (I′), (A′),(I), or (A), or a pharmaceutically acceptable form thereof, incombination with a PI3K-delta selective compound, wherein the PI3K-deltaselective compound is GSK-2269557, GS-9820, GS-1101 (Cal-101), AMG319,or TGR-1202, or a mixture thereof. In one embodiment, the PI3K-deltaselective compound is GS1101. In one embodiment, the cell is a cancercell. In another embodiment, the cell is in a subject. In oneembodiment, the subject is afflicted with a proliferative disease,cancer, autoimmune disease, or inflammatory disease.

In one embodiment, the PI3K-delta selective compound is a compoundselected from US Patent Publication Nos. 20140058103, 20140051699,20140045825, 20140011819, 20130231356, 20130225557, 20120245144,20100305084, 20100256167, 20100168139, 20100152211, and 20100029693. Inone embodiment, the PI3K-delta selective compound is a compound selectedfrom U.S. Pat. Nos. 8,653,077, 8,637,533, 8,623,881, 8,586,597,8,569,296, 8,563,540, 8,492,389, 8,440,651, 8,138,195, 7,932,260, and6,949,535.

For example, a compound provided herein with a delta/gamma selectivityratio of greater than 150 can be combined with a compound that has agamma/delta selectivity ratio of 1000 at various amounts (e.g., a ratioof 10:1 or 40:1 of a gamma selective compound and a delta selectivecompound) to provide synergistic effect in cell lines (e.g., diffuselarge B-cell lymphoma cell lines such as SU-DHL-4, TMD-8 and Farage).

The PI3K-gamma selective compound and PI3K-delta selective compoundcomposition or combination therapy can provide synergistic effect intreating or preventing a PI3K mediated disorder. In one embodiment, thedisorder is a cancer. In one embodiment, the cancer is diffuse largeB-cell lymphoma (e.g., TMD-8 and Farage cell lines), B-cell lymphoma(e.g., karpas-422 cell line), T-cell lymphoma, non-Hodgkin's lymphoma,Hodgkin lymphoma, or anaplastic large cell lymphoma (e.g., HH cellline).

In some embodiments, the synergistic effect can be characterized by anisobologram. Potency shifting is usually shown using an isobologramwhich shows how much less a compound is required in combination toachieve a desired effect level, when compared to the single agent dosesneeded to reach that effect. The choice of effect level for theisobologram display and combination index calculations can either bemanually or automatically selected in the Chalice Analyzer. Potencyshifting is scored as the combination index (CI). Chou et al., AdvEnzyme Regul 1984; 22: 27-55. The CI is a rough estimate of how much acompound was needed in combination relative to the single agent dosesrequired to achieve the chosen effect level, and a value of 0.1 meansthat only a tenth of equivalent amounts of the single agents were neededfor the combination to reach the same effect level. Additive effect isCI=1.0. Synergistic effect is CI<1. Antagonistic effect is CI>1.0.

In some embodiment, the synergistic effect is characterized by SynergyScore.

Different stimuli can be used to preferentially induce T-cell orCLL-cell migration. For example, CCL19 and CCL21 stimuli selectivelyinduce migration of both CLL and T-cells. CXCL13 is CLL-cell specific,whereas CXCL12 is T-cell specific. As such, stimuli CXCL13 and CXCL12can be used to induce CLL-cell and T-cell migrations, respectively. ThePI3K-gamma selective compounds provided herein can inhibitcancer-promoting cell migration, e.g., CXCL 12-induced T-cell migration.In some embodiments, elevated pAKT levels indicate that theCXCL12-induced migration machinery is activated. Consequently, in someembodiments, the PI3K-gamma selective compound, e.g., Compound 1,interferes with AKT signaling and/or reduces pAKT levels in the T-cells.In one embodiment, the PI3K-gamma selective compound is a compound thathas a delta/gamma selectivity ratio of greater than about 50, such asCompound 1. In another embodiment, the PI3K-delta selective compound isa compound that has a gamma/delta selectivity ratio of greater thanabout 50. The gamma selective compound can be more potent than a deltaselective compound at inhibiting cancer-promoting cell migration, e.g.,CXCL12-induced T cell migration in CLL PBMCs. The ability to inhibit themigration of cancer-promoting cells can stop the growth of cancers byblocking the migration of cells that promote cancer growth to the cancercell niche. In another embodiment, gamma or delta selective compoundscan inhibit the migration of cancer cells themselves and limit cancercell dissemination. As such, the gamma selective compounds providedherein can be used to treat and/or prevent cancer, or slow down theprogression of cancer or metastasis. Treatment with a combination ofgamma and delta selective compounds can have an earlier response timecompared to a delta selective compound alone, for example in B cellmeditated cancers.

In one embodiment, a compound provided herein (e.g., Compound 1), or apharmaceutically acceptable form thereof, is administered in combinationwith a chemotherapy (e.g., temozolomide) for the treatment of a cancer.In one embodiment, the cancer is glioblastoma (e.g., glioblastomamultiforme). In one embodiment, provided herein is a method of treatingglioblastoma in a subject, comprising administering to the subject atherapeutically effective amount of Compound 1, or a pharmaceuticallyacceptable form thereof, in combination with a chemotherapy. In oneembodiment, the compound is administered subsequent to the chemotherapy.In one embodiment, the compound is administered concurrently to thechemotherapy. In one embodiment, the compound is administered prior tothe chemotherapy.

In some embodiments, provided herein is a method for using a compoundprovided herein, or a pharmaceutically acceptable form (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein, in combination withradiation therapy in inhibiting abnormal cell growth or treating thehyperproliferative disorder in the subject. Techniques for administeringradiation therapy are known in the art, and these techniques can be usedin the combination therapy described herein. The administration of acompound provided herein in this combination therapy can be determinedas described herein.

In certain embodiments, provided herein are methods of treating a solidtumor in a subject, comprising administering to the subject atherapeutically effective amount of a compound provided herein (e.g.,Compound 1), or a pharmaceutically acceptable form thereof, incombination with a radiation therapy.

In one embodiment, the solid tumor is selected from one or more of: acancer of the pulmonary system, a brain cancer, a cancer of thegastrointestinal tract, a skin cancer, a genitourinary cancer, apancreatic cancer, a lung cancer, a medulloblastoma, a basal cellcarcinoma, a glioma, a breast cancer (e.g., triple negative breastcancer), a prostate cancer, a testicular cancer, an esophageal cancer, ahepatocellular cancer, a gastric cancer, a gastrointestinal stromaltumor (GIST), a colon cancer, a colorectal cancer, an ovarian cancer, amelanoma, a neuroectodermal tumor, head and neck cancer, a sarcoma, asoft-tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, achondrosarcoma, an osteogenic sarcoma, a chordoma, an angiosarcoma, anendotheliosarcoma, a lymphangiosarcoma, a lymphangioendotheliosarcoma, asynovioma, a mesothelioma, a leiomyosarcoma, a cervical cancer, auterine cancer, an endometrial cancer, a carcinoma, a bladder carcinoma,an epithelial carcinoma, a squamous cell carcinoma, an adenocarcinoma, abronchogenic carcinoma, a renal cell carcinoma, a hepatoma, a bile ductcarcinoma, a neuroendocrine cancer, a carcinoid tumor, diffuse typegiant cell tumor, and glioblastoma.

In one embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered after the radiation therapy is administered. Inone embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered at the same time that radiation therapy isadministered. In one embodiment, the compound, or a pharmaceuticallyacceptable form thereof, is administered alone after discontinuing theradiation therapy.

Radiation therapy can be administered through one of several methods, ora combination of methods, including without limitation, external-beamtherapy, internal radiation therapy, implant radiation, stereotacticradiosurgery, systemic radiation therapy, radiotherapy and permanent ortemporary interstitial brachytherapy. The term “brachytherapy,” as usedherein, refers to radiation therapy delivered by a spatially confinedradioactive material inserted into the body at or near a tumor or otherproliferative tissue disease site. The term is intended withoutlimitation to include exposure to radioactive isotopes (e.g., At-211,I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, andradioactive isotopes of Lu). Suitable radiation sources for use as acell conditioner as provided herein include both solids and liquids. Byway of non-limiting example, the radiation source can be a radionuclide,such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solidsource, or other radionuclides that emit photons, beta particles, gammaradiation, or other therapeutic rays. The radioactive material can alsobe a fluid made from any solution of radionuclide(s), e.g., a solutionof I-125 or I-131, or a radioactive fluid can be produced using a slurryof a suitable fluid containing small particles of solid radionuclides,such as Au-198, Y-90. Moreover, the radionuclide(s) can be embodied in agel or radioactive micro spheres.

Without being limited by any theory, a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, or a pharmaceutical composition as providedherein, can render abnormal cells more sensitive to treatment withradiation for purposes of killing and/or inhibiting the growth of suchcells. Accordingly, provided herein is a method for sensitizing abnormalcells in a subject to treatment with radiation which comprisesadministering to the subject an amount of a compound provided herein, ora pharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, which amount is effective in sensitizing abnormalcells to treatment with radiation. The amount of the compound used inthis method can be determined according to the means for ascertainingeffective amounts of such compounds described herein.

In one embodiment, a compound as provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein, can be usedin combination with an amount of one or more substances selected fromanti-angiogenesis agents, signal transduction inhibitors, andantiproliferative agents, glycolysis inhibitors, or autophagyinhibitors.

Other therapeutic agents, such as MMP-2 (matrix-metalloproteinase 2)inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-11(cyclooxygenase 11) inhibitors, can be used in conjunction with acompound provided herein, or a pharmaceutically acceptable form thereof,or a pharmaceutical composition described herein. Such therapeuticagents include, for example, rapamycin, temsirolimus (CCI-779),everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples ofuseful COX-II inhibitors include CELEBREX™ (alecoxib), valdecoxib, androfecoxib. Examples of useful matrix metalloproteinase inhibitors aredescribed in WO 96/33172 (published Oct. 24, 1996), WO 96/27583(published Mar. 7, 1996), European Patent Application No. 97304971.1(filed Jul. 8, 1997), European Patent Application No. 99308617.2 (filedOct. 29, 1999), WO 98/07697 (published Feb. 26, 1998), WO 98/03516(published Jan. 29, 1998), WO 98/34918 (published Aug. 13, 1998), WO98/34915 (published Aug. 13, 1998), WO 98/33768 (published Aug. 6,1998), WO 98/30566 (published Jul. 16, 1998), European PatentPublication 606,046 (published Jul. 13, 1994), European PatentPublication 931, 788 (published Jul. 28, 1999), WO 90/05719 (publishedMay 31, 1990), WO 99/52910 (published Oct. 21, 1999), WO 99/52889(published Oct. 21, 1999), WO 99/29667 (published Jun. 17, 1999), PCTInternational Application No. PCT/IB98/01113 (filed Jul. 21, 1998),European Patent Application No. 99302232.1 (filed Mar. 25, 1999), GreatBritain Patent Application No. 9912961.1 (filed Jun. 3, 1999), U.S.Provisional Application No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat.No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issuedJan. 19, 1999), and European Patent Publication 780,386 (published Jun.25, 1997), all of which are incorporated herein in their entireties byreference. In some embodiments, MMP-2 and MMP-9 inhibitors are thosethat have little or no activity inhibiting MMP-1. Other embodimentsinclude those that selectively inhibit MMP-2 and/or AMP-9 relative tothe other matrix-metalloproteinases (e.g., MAP-1, MMP-3, MMP-4, MMP-5,MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Somenon-limiting examples of MMP inhibitors are AG-3340, RO 32-3555, and RS13-0830.

Autophagy inhibitors include, but are not limited to, chloroquine,3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1,5-amino-4-imidazole carboxamide riboside (AICAR), okadaic acid,autophagy-suppressive algal toxins which inhibit protein phosphatases oftype 2A or type 1, analogues of cAMP, and drugs which elevate cAMPlevels such as adenosine, LY204002, N6-mercaptopurine riboside, andvinblastine. In addition, antisense or siRNAs that inhibit expression ofproteins including, but not limited to ATG5 (which are implicated inautophagy), can also be used.

In some embodiments, provided herein is a method of and/or apharmaceutical composition for treating a cardiovascular disease in asubject which comprises an amount of a compound provided herein, or apharmaceutically acceptable form (e.g., pharmaceutically acceptablesalts, hydrates, solvates, isomers, prodrugs, and isotopically labeledderivatives) thereof, and an amount of one or more therapeutic agentsuse for the treatment of cardiovascular diseases.

Exemplary agents for use in cardiovascular disease applications areanti-thrombotic agents, e.g., prostacyclin and salicylates, thrombolyticagents, e.g., streptokinase, urokinase, tissue plasminogen activator(TPA) and anisoylated plasminogen-streptokinase activator complex(APSAC), anti-platelets agents, e.g., acetylsalicylic acid (ASA) andclopidrogel, vasodilating agents, e.g., nitrates, calcium channelblocking drugs, antiproliferative agents, e.g., colchicine andalkylating agents, intercalating agents, growth modulating factors suchas interleukins, transformation growth factor-beta and congeners ofplatelet derived growth factor, monoclonal antibodies directed againstgrowth factors, anti-inflammatory agents, both steroidal andnon-steroidal, and other agents that can modulate vessel tone, function,arteriosclerosis, and the healing response to vessel or organ injurypost intervention. Antibiotics can also be included in combinations orcoatings. Moreover, a coating can be used to effect therapeutic deliveryfocally within the vessel wall. By incorporation of the active agent ina swellable polymer, the active agent will be released upon swelling ofthe polymer.

In one embodiment, a compound provided herein, or a pharmaceuticallyacceptable form (e.g., pharmaceutically acceptable salts, hydrates,solvates, isomers, prodrugs, and isotopically labeled derivatives)thereof, or a pharmaceutical composition as provided herein, can beformulated or administered in conjunction with liquid or solid tissuebarriers also known as lubricants. Examples of tissue barriers include,but are not limited to, polysaccharides, polyglycans, seprafilm,interceed and hyaluronic acid.

Medicaments which can be administered in conjunction with a compoundprovided herein, or a pharmaceutically acceptable form (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, include anysuitable drugs usefully delivered by inhalation for example, analgesics,e.g., codeine, dihydromorphine, ergotamine, fentanyl or morphine;anginal preparations, e.g., diltiazem; antiallergics, e.g. cromoglycate,ketotifen or nedocromil; anti-infectives, e.g., cephalosporins,penicillins, streptomycin, sulphonamides, tetracyclines or pentamidine;antihistamines, e.g., methapyrilene; anti-inflammatories, e.g.,beclomethasone, flunisolide, budesonide, tipredane, triamcinoloneacetonide or fluticasone; antitussives, e.g., noscapine;bronchodilators, e.g., ephedrine, adrenaline, fenoterol, formoterol,isoprenaline, metaproterenol, phenylephrine, phenylpropanolamine,pirbuterol, reproterol, rimiterol, salbutamol, salmeterol, terbutalin,isoetharine, tulobuterol, orciprenaline or(−)-4-amino-3,5-dichloro-α[[[6[2-(2-pyridinyl)ethoxy]hexyl]-amino]methyl]benzenemethanol;diuretics, e.g., amiloride; anticholinergics e.g., ipratropium, atropineor oxitropium; hormones, e.g., cortisone, hydrocortisone orprednisolone; xanthines e.g., aminophylline, choline theophyllinate,lysine theophyllinate or theophylline; and therapeutic proteins andpeptides, e.g., insulin or glucagon. It will be clear to a personskilled in the art that, where appropriate, the medicaments can be usedin the form of salts (e.g., as alkali metal or amine salts or as acidaddition salts) or as esters (e.g., lower alkyl esters) to optimize theactivity and/or stability of the medicament.

Other exemplary therapeutic agents useful for a combination therapyinclude, but are not limited to, agents as described above, radiationtherapy, hormone antagonists, hormones and their releasing factors,thyroid and antithyroid drugs, estrogens and progestins, androgens,adrenocorticotropic hormone; adrenocortical steroids and their syntheticanalogs; inhibitors of the synthesis and actions of adrenocorticalhormones, insulin, oral hypoglycemic agents, and the pharmacology of theendocrine pancreas, agents affecting calcification and bone turnover:calcium, phosphate, parathyroid hormone, vitamin D, calcitonin, vitaminssuch as water-soluble vitamins, vitamin B complex, ascorbic acid,fat-soluble vitamins, vitamins A, K, and E, growth factors, cytokines,chemokines, muscarinic receptor agonists and antagonists;anticholinesterase agents; agents acting at the neuromuscular junctionand/or autonomic ganglia; catecholamines, sympathomimetic drugs, andadrenergic receptor agonists or antagonists; and 5-hydroxytryptamine(5-HT, serotonin) receptor agonists and antagonists.

Therapeutic agents can also include agents for pain and inflammationsuch as histamine and histamine antagonists, bradykinin and bradykininantagonists, 5-hydroxytryptamine (serotonin), lipid substances that aregenerated by biotransformation of the products of the selectivehydrolysis of membrane phospholipids, eicosanoids, prostaglandins,thromboxanes, leukotrienes, aspirin, nonsteroidal anti-inflammatoryagents, analgesic-antipyretic agents, agents that inhibit the synthesisof prostaglandins and thromboxanes, selective inhibitors of theinducible cyclooxygenase, selective inhibitors of the induciblecyclooxygenase-2, autacoids, paracrine hormones, somatostatin, gastrin,cytokines that mediate interactions involved in humoral and cellularimmune responses, lipid-derived autacoids, eicosanoids, β-adrenergicagonists, ipratropium, glucocorticoids, methylxanthines, sodium channelblockers, opioid receptor agonists, calcium channel blockers, membranestabilizers and leukotriene inhibitors.

Additional therapeutic agents contemplated herein include diuretics,vasopressin, agents affecting the renal conservation of water, rennin,angiotensin, agents useful in the treatment of myocardial ischemia,anti-hypertensive agents, angiotensin converting enzyme inhibitors,β-adrenergic receptor antagonists, agents for the treatment ofhypercholesterolemia, and agents for the treatment of dyslipidemia.

Other therapeutic agents contemplated herein include drugs used forcontrol of gastric acidity, agents for the treatment of peptic ulcers,agents for the treatment of gastroesophageal reflux disease, prokineticagents, antiemetics, agents used in irritable bowel syndrome, agentsused for diarrhea, agents used for constipation, agents used forinflammatory bowel disease, agents used for biliary disease, agents usedfor pancreatic disease. Therapeutic agents include, but are not limitedto, those used to treat protozoan infections, drugs used to treatMalaria, Amebiasis, Giardiasis, Trichomoniasis, Trypanosomiasis, and/orLeishmaniasis, and/or drugs used in the chemotherapy of helminthiasis.Other therapeutic agents include, but are not limited to, antimicrobialagents, sulfonamides, trimethoprim-sulfamethoxazole quinolones, andagents for urinary tract infections, penicillins, cephalosporins, andother, β-Lactam antibiotics, an agent containing an aminoglycoside,protein synthesis inhibitors, drugs used in the chemotherapy oftuberculosis, mycobacterium avium complex disease, and leprosy,antifungal agents, antiviral agents including nonretroviral agents andantiretroviral agents.

Examples of therapeutic antibodies that can be combined with a compoundprovided herein include but are not limited to anti-receptor tyrosinekinase antibodies (cetuximab, panitumumab, trastuzumab), anti CD20antibodies (rituximab, tositumomab), and other antibodies such asalemtuzumab, bevacizumab, and gemtuzumab.

Moreover, therapeutic agents used for immuno-modulation, such asimmuno-modulators, immuno-suppressive agents, tolerogens, andimmunostimulants are contemplated by the methods herein. In addition,therapeutic agents acting on the blood and the blood-forming organs,hematopoietic agents, growth factors, minerals, and vitamins,anticoagulant, thrombolytic, and anti-platelet drugs are alsocontemplated by the methods herein.

In exemplary embodiments, for treating renal carcinoma, one can combinea compound provided herein, or a pharmaceutically acceptable form (e.g.,pharmaceutically acceptable salts, hydrates, solvates, isomers,prodrugs, and isotopically labeled derivatives) thereof, or apharmaceutical composition as provided herein, with sorafenib and/oravastin. For treating an endometrial disorder, one can combine acompound provided herein with doxorubincin, taxotere (taxol), and/orcisplatin (carboplatin). For treating ovarian cancer, one can combine acompound provided herein with cisplatin, carboplatin, docetaxel,doxorubincin, topotecan, and/or tamoxifen. For treating breast cancer,one can combine a compound provided herein with paclitaxel or docetaxel,gemcitabine, capecitabine, tamoxifen, letrozole, erlotinib, lapatinib,PD0325901, bevacizumab, trastuzumab, OSI-906, and/or OSI-930. Fortreating lung cancer, one can combine a compound as provided herein withpaclitaxel, docetaxel, gemcitabine, cisplatin, pemetrexed, erlotinib,PD0325901, and/or bevacizumab.

In some embodiments, the disorder to be treated, prevented and/ormanaged is a hematological cancer, e.g., lymphoma (e.g., T-celllymphoma; NHL), myeloma (e.g., multiple myeloma), and leukemia (e.g.,CLL), and a compound provided herein is used in combination with: HDACinhibitors such as vorinostat, romidepsin and ACY-1215; mTOR inhibitorssuch as everolimus; anti-folates such as pralatrexate; nitrogen mustardsuch as bendamustine; gemcitabine, optionally in further combinationwith oxaliplatin; rituximab-cyclophosphamide combination; PI3Kinhibitors such as RP-6530, TG 100-115, RV1729, GS-1101, XL 499,GDC-0941, and AMG-319; angiogenesis inhibitors such as pomalidomide orBTK inhibitors such as ibrutinib, AVL-292, Dasatinib, LFM-AI3,ONO-WG-307, and GDC-0834. In some embodiments, the disorder to betreated, prevented and/or managed is DLBCL, and a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, is used in combination with HDACinhibitors provided herein. In one particular embodiment, the HDACinhibitor is ACY-1215.

In some embodiments, the disorder to be treated, prevented and/ormanaged is DLBCL, and a compound provided herein (e.g., compound 1), ora pharmaceutically acceptable derivative (e.g., salt or solvate)thereof, is used in combination with BTK inhibitors provided herein. Inone particular embodiment, the BTK inhibitor is ibrutinib. In oneembodiment, the BTK inhibitor is AVL-292.

In some embodiments, the disorder to be treated, prevented and/ormanaged is DLBCL, and a compound provided herein (e.g., compound 1), ora pharmaceutically acceptable derivative (e.g., salt or solvate)thereof, is used in combination with IRAK inhibitors provided herein. Inone particular embodiment, the IRAK4 inhibitor is ND-2110 or ND-2158.

In some embodiments, the disorder to be treated, prevented and/ormanaged is WM, and a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with BTK inhibitors provided herein. In oneparticular embodiment, the BTK inhibitor is ibrutinib. In oneembodiment, the BTK inhibitor is AVL-292.

In some embodiments, the disorder to be treated, prevented and/ormanaged is WM, and a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with IRAK4 inhibitors provided herein. In oneparticular embodiment, the IRAK4 inhibitor is ND-2110 or ND-2158.

In some embodiments, the disorder to be treated, prevented and/ormanaged is T-ALL, the subject/patient has a PTEN deficiency, and acompound provided herein (e.g., compound 1), or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, is used incombination with doxorubicin and/or vincristine.

In certain embodiments, wherein inflammation (e.g., arthritis, asthma)is treated, prevented and/or managed, a compound provided herein can becombined with, for example: PI3K inhibitors such as RP-6530, TG 100-115,RV1729, GS-1101, XL 499, GDC-0941, and AMG-319; BTK inhibitors such asibrutinib and AVL-292; JAK inhibitors such as tofacitinib and GLPG0636;SYK inhibitors such as fostamatinib.

In certain embodiments wherein asthma is treated, prevented and/ormanaged, a compound provided herein can be combined with, for example:beta 2-agonists such as, but not limited to, albuterol (Proventil®, orVentolin®), salmeterol (Serevent®), formoterol (Foradil®),metaproterenol (Alupent®), pirbuterol (MaxAir®), and terbutalinesulfate; corticosteroids such as, but not limited to, budesonide (e.g.,Pulmicort®), flunisolide (e.g., AeroBid Oral Aerosol Inhaler® orNasalide Nasal Aerosol®), fluticasone (e.g., Flonase® or Flovent®) andtriamcinolone (e.g., Azmacort®); mast cell stabilizers such as cromolynsodium (e.g., Intal® or Nasalcrom®) and nedocromil (e.g., Tilade®);xanthine derivatives such as, but not limited to, theophylline (e.g.,Aminophyllin®, Theo-24® or Theolair®); leukotriene receptor antagonistssuch as, but are not limited to, zafirlukast (Accolate®), montelukast(Singulair®), and zileuton (Zyflo®); and adrenergic agonists such as,but are not limited to, epinephrine (Adrenalin®, Bronitin®, EpiPen® orPrimatene Mist®).

In certain embodiments wherein arthritis is treated, prevented and/ormanaged, a compound provided herein can be combined with, for example:TNF antagonist (e.g., a TNF antibody or fragment, a soluble TNF receptoror fragment, fusion proteins thereof, or a small molecule TNFantagonist); other biologic antirhheumatics (e.g., IL-6 antagonists,IL-1 antagonists, costimulatory modulators); an antirheumatic (e.g.,methotrexate, auranofin, aurothioglucose, azathioprine, etanercept, goldsodium thiomalate, chrloroquine, hydroxychloroquine sulfate,leflunomide, sulfasalzine, penicillamine); a muscle relaxant; anarcotic; a non-steroid anti-inflammatory drug (NSAID); an analgesic; ananesthetic; a sedative; a local anesthetic; a neuromuscular blocker; anantimicrobial (e.g., an aminoglycoside, an antifungal, an antiparasitic,an antiviral, a carbapenem, cephalosporin, a fluoroquinolone, amacrolide, a penicillin, a sulfonamide, a tetracycline, anotherantimicrobial); an antipsoriatic; a corticosteroid; an anabolic steroid;a cytokine or a cytokine antagonist; a calcineurin inhibitor (e.g.,cyclosporine, tacrolimus).

In some embodiments, a compound provided herein (e.g., compound 1), oran enantiomer or a mixture of enantiomers thereof, or a pharmaceuticallyacceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorphthereof) is administered in combination with an agent for the treatmentof rheumatoid arthritis. Examples of agents for the treatment ofrheumatoid arthritis include, but are not limited to, various NSAIDs,corticosteroids, sulfasalazine, auranofin, methotrexate, azathioprine,penicillamine, cyclosporine, Arava (leflunomide), TNF inhibitors (e.g.,Enbrel (etanercept), Remicade (infliximab), Humira (adalimumab), Simponi(golimumab), and Cimzia (certolizumab)), IL-1 inhibitors (e.g., Kineret(anakinra)), T-cell costimulatory modulators (e.g., Orencia(abatacept)), Anti-CD20 (e.g., Rituxan (rituximab)), and IL-6 inhibitors(e.g., Actemra (tocilizumab)). In one embodiment, the agent is Cimzia(certolizumab). In another embodiment, the agent is Actemra(tocilizumab).

In some embodiments, a compound provided herein (e.g., compound 1), oran enantiomer or a mixture of enantiomers thereof, or a pharmaceuticallyacceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorphthereof) is administered in combination with an agent for rheumatology.Examples of agents for rheumatology include, but are not limited to,Rayos (prednisone), Stendra (avanafil), Actemra (tocilizumab), Duexis(ibuprofen and famotidine), Actemra (tocilizumab), Krystexxa(pegloticase), Vimovo (naproxen+esomeprazole), Cimzia (certolizumabpegol), Colcrys (colchicine), Pennsaid (diclofenac sodium topicalsolution), Simponi (golimumab), Uloric (febuxostat), Orencia(abatacept), Elaprase (idursulfase), Orencia (abatacept), Vioxx(rofecoxib), Enbrel (etanercept), Humira (adalimumab), Remicade(infliximab), Bextra, Kineret, Remicade (infliximab), Supartz, Mobic(meloxicam), Vivelle (estradiol transdermal system), Lodine XL(etodolac), Arava, Salagen, Arthrotec, Etodolac, Ketoprofen, Synvisc,Tolmetin Sodium, Azulfidine EN-tabs Tablets (sulfasalazine delayedrelease tablets, USP), and Naprelan (naproxen sodium).

In some embodiments, the second agent is selected from belimumab,AGS-009, rontalizumab, vitamin D3, sifalimumab, AMG 811, IFNα Kinoid,CEP33457, epratuzumab, LY2127399, Ocrelizumab, Atacicept, A-623,SBI-087, AMG557, laquinimod, rapamycin, cyclophosphamide, azathioprine,mycophenolate, leflunomide, methotrexate, CNTO 136, tamibarotene,N-acetylcysteine, CDP7657, hydroxychloroquine, rituximab, carfilzomib,bortezomib, ONX 0914, IMO-3100, DV1179, sulfasalazine, and chloroquine.In one embodiment, the second agent is methotrexate, sulfasalazine,chloroquine, or hydroxychloroquine. In one embodiment, the second agentis methotrexate.

In certain embodiments wherein psoriasis is treated, prevented and/ormanaged, a compound provided herein can be combined with, for example:budesonide, epidermal growth factor, corticosteroids, cyclosporine,sulfasalazine, aminosalicylates, 6-mercaptopurine, azathioprine,metronidazole, lipoxygenase inhibitors, mesalamine, olsalazine,balsalazide, antioxidants, thromboxane inhibitors, IL-1 receptorantagonists, anti-IL-1β monoclonal antibodies, anti-IL-6 monoclonalantibodies, growth factors, elastase inhibitors, pyridinyl-imidazolecompounds, antibodies or agonists of TNF, LT, IL-1, IL-2, IL-6, IL-7,IL-8, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF, and PDGF, antibodies ofCD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD90 or theirligands, methotrexate, cyclosporine, FK506, rapamycin, mycophenolatemofetil, leflunomide, NSAIDs, ibuprofen, corticosteroids, prednisolone,phosphodiesterase inhibitors, adenosine agonists, antithrombotic agents,complement inhibitors, adrenergic agents, IRAK, NIK, IKK, p38, MAPkinase inhibitors, IL-1β converting enzyme inhibitors, TNFα convertingenzyme inhibitors, T-cell signaling inhibitors, metalloproteinaseinhibitors, sulfasalazine, azathioprine, 6-mercaptopurines, angiotensinconverting enzyme inhibitors, soluble cytokine receptors, soluble p55TNF receptor, soluble p75 TNF receptor, sIL-1RI, sIL-1RII, sIL-6R,anti-inflammatory cytokines, IL-4, IL-10, IL-11, IL-13 and TGFβ.

In certain embodiments wherein fibrosis or fibrotic condition of thebone marrow is treated, prevented and/or managed, a compound providedherein can be combined with, for example, a Jak2 inhibitor (including,but not limited to, INCB018424, XL019, TG101348, or TG101209), animmuno-modulator, e.g., an IMID® (including, but not limited tothalidomide, lenalidomide, or panolinomide), hydroxyurea, an androgen,erythropoietic stimulating agents, prednisone, danazol, HDAC inhibitors,or other agents or therapeutic modalities (e.g., stem cell transplants,or radiation).

In certain embodiments wherein fibrosis or fibrotic condition of theheart is treated, prevented and/or managed, a compound provided hereincan be combined with, for example, eplerenone, furosemide, pycnogenol,spironolactone, TcNC100692, torasemide (e.g., prolonged release form oftorasemide), or combinations thereof.

In certain embodiments wherein fibrosis or fibrotic condition of thekidney is treated, prevented and/or managed, a compound provided hereincan be combined with, for example, cyclosporine, cyclosporine A,daclizumab, everolimus, gadofoveset trisodium (ABLAVAR®), imatinibmesylate (GLEEVEC®), matinib mesylate, methotrexate, mycophenolatemofetil, prednisone, sirolimus, spironolactone, STX-100, tamoxifen,TheraCLEC™, or combinations thereof.

In certain embodiments wherein fibrosis or fibrotic condition of theskin is treated, prevented and/or managed, a compound provided hereincan be combined with, for example, Bosentan (Tracleer), p144,pentoxifylline; pirfenidone; pravastatin, STI571, Vitamin E, orcombinations thereof.

In certain embodiments wherein fibrosis or fibrotic condition of thegastrointestinal system is treated, prevented and/or managed, a compoundprovided herein can be combined with, for example, ALTU-135, bucelipasealfa (INN), DCI1020, EUR-1008 (ZENPEP™), ibuprofen, Lym-X-Sorb powder,pancrease MT, pancrelipase (e.g., pancrelipase delayed release), pentadecanoic acid (PA), repaglinide, TheraCLEC™, triheptadecanoin (THA),ULTRASE MT20, ursodiol, or combinations thereof.

In certain embodiments wherein fibrosis or fibrotic condition of thelung is treated, prevented and/or managed, a compound provided hereincan be combined with, for example, 18-FDG, AB0024, ACT-064992(macitentan), aerosol interferon-gamma, aerosolized human plasma-derivedalpha-1 antitrypsin, alpha1-proteinase inhibitor, ambrisentan, amikacin,amiloride, amitriptyline, anti-pseudomonas IgY gargle, ARIKACE™,AUREXIS® (tefibazumab), AZAPRED, azathioprine, azithromycin,azithromycin, AZLI, aztreonam lysine, BIBF1120, Bio-25 probiotic,bosentan, Bramitob®, calfactant aerosol, captopril, CC-930, ceftazidime,ceftazidime, cholecalciferol (Vitamin D3), ciprofloxacin (CIPRO®,BAYQ3939), CNTO 888, colistin CF, combined Plasma Exchange (PEX),rituximab, and corticosteroids, cyclophosphamide, dapsone, dasatinib,denufosol tetrasodium (INS37217), dornase alfa (PULMOZYME®), EPI-hNE4,erythromycin, etanercept, FG-3019, fluticasone, FTI, GC1008, GS-9411,hypertonic saline, ibuprofen, iloprost inhalation, imatinib mesylate(GLEEVEC®), inhaled sodium bicarbonate, inhaled sodium pyruvate,interferon gamma-1b, interferon-alpha lozenges, isotonic saline, IW001,KB001, losartan, lucinactant, mannitol, meropenem, meropenem infusion,miglustat, minocycline, Moli1901, MP-376 (levofloxacin solution forinhalation), mucoid exopolysaccharide P. aeruginosa immune globulin IV,mycophenolate mofetil, n-acetylcysteine, N-acetylcysteine (NAC), NaCl6%, nitric oxide for inhalation, obramycin, octreotide, oligoG CF-5/20,Omalizumab, pioglitazone, piperacillin-tazobactam, pirfenidone,pomalidomide (CC-4047), prednisone, prevastatin, PRM-151, QAX576,rhDNAse, SB656933, SB-656933-AAA, sildenafil, tamoxifen, technetium[Tc-99m] sulfur colloid and Indium [In-111] DTPA, tetrathiomolybdate,thalidomide, ticarcillin-clavulanate, tiotropium bromide, tiotropiumRESPIMAT® inhaler, tobramycin (GERNEBCIN®), treprostinil, uridine,valganciclovir (VALCYTE®), vardenafil, vitamin D3, xylitol, zileuton, orcombinations thereof.

In certain embodiments wherein fibrosis or fibrotic condition of theliver is treated, prevented and/or managed, a compound provided hereincan be combined with, for example, adefovir dipivoxil, candesartan,colchicine, combined ATG, mycophenolate mofetil, and tacrolimus,combined cyclosporine microemulsion and tacrolimus, elastometry,everolimus, FG-3019, Fuzheng Huayu, GI262570, glycyrrhizin (monoammoniumglycyrrhizinate, glycine, L-cysteine monohydrochloride), interferongamma-1b, irbesartan, losartan, oltipraz, ORAL IMPACT®, peginterferonalfa-2a, combined peginterferon alfa-2a and ribavirin, peginterferonalfa-2b (SCH 54031), combined peginterferon alpha-2b and ribavirin,praziquantel, prazosin, raltegravir, ribavirin (REBETOL®, SCH 18908),ritonavir-boosted protease inhibitor, pentoxyphilline, tacrolimus,tauroursodeoxycholic acid, tocopherol, ursodiol, warfarin, orcombinations thereof.

In certain embodiments wherein cystic fibrosis is treated, preventedand/or managed, a compound provided herein can be combined with, forexample, 552-02, 5-methyltetrahydrofolate and vitamin B12, Ad5-CB-CFTR,Adeno-associated virus-CFTR vector, albuterol, alendronate, alphatocopherol plus ascorbic acid, amiloride HCl, aquADEK™, ataluren(PTC124), AZD1236, AZD9668, azithromycin, bevacizumab, biaxin(clarithromycin), BIIL 283 BS (amelubent), buprofen, calcium carbonate,ceftazidime, cholecalciferol, choline supplementation, CPX, cysticfibrosis transmembrane conductance regulator, DHA-rich supplement,digitoxin, cocosahexaenoic acid (DHA), doxycycline, ECGC, ecombinanthuman IGF-1, educed glutathione sodium salt, ergocalciferol (vitaminD2), fluorometholone, gadobutrol (GADOVIST®, BAY86-4875), gentamicin,ghrelin, glargine, glutamine, growth hormone, GS-9411, H5.001CBCFTR,human recombinant growth hormone, hydroxychloroquine, hyperbaric oxygen,hypertonic saline, IH636 grape seed proanthocyanidin extract, insulin,interferon gamma-1b, IoGen (molecular iodine), iosartan potassium,isotonic saline, itraconazole, IV gallium nitrate (GANITE®) infusion,ketorolac acetate, lansoprazole, L-arginine, linezolid, lubiprostone,meropenem, miglustat, MP-376 (levofloxacin solution for inhalation),normal saline IV, Nutropin AQ, omega-3 triglycerides, pGM169/GL67A,pGT-1 gene lipid complex, pioglitazone, PTC124, QAU145, salmeterol,SB656933, SB656933, simvastatin, sitagliptin, sodium 4-phenylbutyrate,standardized turmeric root extract, tgAAVCF, TNF blocker, TOBI,tobramycin, tocotrienol, unconjugated Isoflavones 100, vitamin: cholinebitartrate (2-hydroxyethyl) trimethylammonium salt 1:1, VX-770, VX-809,Zinc acetate, or combinations thereof.

In some embodiments, a compound provided herein is administered incombination with an agent that inhibits IgE production or activity. Insome embodiments, the PI3K inhibitor (e.g., PI3Kδ inhibitor) isadministered in combination with an inhibitor of mTOR. Agents thatinhibit IgE production are known in the art and they include but are notlimited to one or more of TEI-9874,2-(4-(6-cyclohexyloxy-2-naphtyloxy)phenylacetamide)benzoic acid,rapamycin, rapamycin analogs (i.e. rapalogs), TORC1 inhibitors, TORC2inhibitors, and any other compounds that inhibit mTORC1 and mTORC2.Agents that inhibit IgE activity include, for example, anti-IgEantibodies such as for example Omalizumab and TNX-901.

In certain embodiments wherein scleroderma is treated, prevented and/ormanaged, a compound provided herein can be combined with, for example:an immunosuppressant (e.g., methotrexate, azathioprine (Imuran®),cyclosporine, mycophenolate mofetil (Cellcept®), and cyclophosphamide(Cytoxan®)); T-cell-directed therapy (e.g., halofuginone, basiliximab,alemtuzumab, abatacept, rapamycin); B-cell directed therapy (e.g.,rituximab); autologous hematopoietic stem cell transplantation; achemokine ligand receptor antagonist (e.g., an agent that targets theCXCL12/CSCR4 axis (e.g., AMD3100)); a DNA methylation inhibitor (e.g.,5-azacytidine); a histone deacetylase inhibitor (e.g., trichostatin A);a statin (e.g., atorvastatin, simvastatin, pravastatin); an endothelinreceptor antagonist (e.g., Bosentan®); a phosphodiesterase type Vinhibitor (e.g., Sildenafil®); a prostacyclin analog (e.g.,trepostinil); an inhibitor of cytokine synthesis and/or signaling (e.g.,Imatinib mesylate, Rosiglitazone, rapamycin, antitransforming growthfactor β1 (anti-TGFβ1) antibody, mycophenolate mofetil, an anti-IL-6antibody (e.g., tocilizumab)); corticosteroids; nonsteroidalanti-inflammatory drugs; light therapy; and blood pressure medications(e.g., ACE inhibitors).

In certain embodiments wherein inflammatory myopathies are treated,prevented and/or managed, a compound provided herein can be combinedwith, for example: topical creams or ointments (e.g., topicalcorticosteroids, tacrolimus, pimecrolimus); cyclosporine (e.g., topicalcyclosporine); an anti-interferon therapy, e.g., AGS-009, Rontalizumab(rhuMAb IFNalpha), Vitamin D3, Sifalimumab (MEDI-545), AMG 811, IFNαKinoid, or CEP33457. In some embodiments, the other therapy is an IFN-αtherapy, e.g., AGS-009, Rontalizumab, Vitamin D3, Sifalimumab (MEDI-545)or IFNα Kinoid; corticosteroids such as prednisone (e.g., oralprednisone); immunosuppressive therapies such as methotrexate (Trexall®,Methotrexate®, Rheumatrex®), azathioprine (Azasan®, Imuran®),intravenous immunoglobulin, tacrolimus (Prograf®), pimecrolimus,cyclophosphamide (Cytoxan®), and cyclosporine (Gengraf®, Neoral®,Sandimmune®); anti-malarial agents such as hydroxychloroquine(Plaquenil®) and chloroquine (Aralen®); total body irradiation;rituximab (Rituxan®); TNF inhibitors (e.g., etanercept (Enbrel®),infliximab (Remicade®)); AGS-009; Rontalizumab (rhuMAb IFNalpha);Vitamin D3; Sifalimumab (MEDI-545); AMG 811; IFNα Kinoid; CEP33457;agents that inhibit IgE production such as TEI-9874,2-(4-(6-cyclohexyloxy-2-naphtyloxy)phenylacetamide)benzoic acid,rapamycin, rapamycin analogs (i.e. rapalogs), TORC1 inhibitors, TORC2inhibitors, and any other compounds that inhibit mTORC1 and mTORC2;agents that inhibit IgE activity such as anti-IgE antibodies (e.g.,Omalizumab and TNX-90); and additional therapies such as physicaltherapy, exercise, rest, speech therapy, sun avoidance, heat therapy,and surgery.

In certain embodiments wherein myositis (e.g., dermatomysitis) istreated, prevented and/or managed, a compound provided herein can becombined with, for example: corticosteroids; corticosteroid sparingagents such as, but not limited to, azathioprine and methotrexate;intravenous immunoglobulin; immunosuppressive agents such as, but notlimited to, tacrolimus, cyclophosphamide and cyclosporine; rituximab;TNFα inhibitors such as, but not limited to, etanercept and infliximab;growth hormone; growth hormone secretagogues such as, but not limitedto, MK-0677, L-162752, L-163022, NN703 ipamorelin, hexarelin, GPA-748(KP102, GHRP-2), and LY444711 (Eli Lilly); other growth hormone releasestimulators such as, but not limited to, Geref, GHRH (1-44), Somatorelin(GRF 1-44), ThGRF genotropin, L-DOPA, glucagon, and vasopressin; andinsulin-like growth factor.

In certain embodiments wherein Sjögren's syndrome is treated, preventedand/or managed, a compound provided herein can be combined with, forexample: pilocarpine; cevimeline; nonsteroidal anti-inflammatory drugs;arthritis medications; antifungal agents; cyclosporine;hydroxychloroquine; prednisone; azathioprine; and cyclophamide.

Administration of a compound provided herein, or a pharmaceuticallyacceptable form thereof, can be effected by any method that enablesdelivery of the compound to the site of action. An effective amount of acompound provided herein, or a pharmaceutically acceptable form thereof,can be administered in either single or multiple doses by any of theaccepted modes of administration of agents having similar utilities,including rectal, buccal, intranasal, and transdermal routes, byintra-arterial injection, intravenously, intraperitoneally,parenterally, intramuscularly, subcutaneously, orally, topically, as aninhalant, or via an impregnated or coated device such as a stent, forexample, or an artery-inserted cylindrical polymer.

When a compound provided herein, or a pharmaceutically acceptable formthereof, is administered in a pharmaceutical composition that comprisesone or more agents, and the agent has a shorter half-life than thecompound provided herein, unit dose forms of the agent and the compoundas provided herein can be adjusted accordingly.

In some embodiments, the compound provided herein and the second agentare administered as separate compositions, e.g., pharmaceuticalcompositions. In some embodiments, the PI3K modulator and the agent areadministered separately, but via the same route (e.g., both orally orboth intravenously). In other embodiments, the PI3K modulator and theagent are administered in the same composition, e.g., pharmaceuticalcomposition.

In some embodiments, a compound provided herein (e.g., compound 1), oran enantiomer or a mixture of enantiomers thereof, or a pharmaceuticallyacceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorphthereof) is administered in combination with an agent for pulmonary orrespiratory diseases. Examples of agents for pulmonary or respiratorydiseases include, but are not limited to, Dymista (azelastinehydrochloride and fluticasone propionate), Kalydeco (ivacaftor), Qnasl(beclomethasone dipropionate) nasal aerosol, Rayos (prednisone)delayed-release tablets, Surfaxin (lucinactant), Tudorza Pressair(aclidinium bromide inhalation powder), Arcapta (indacaterol maleateinhalation powder), Daliresp (roflumilast), Xalkori (crizotinib),Cayston (aztreonam for inhalation solution), Dulera (mometasonefuroate+formoterol fumarate dihydrate), Teflaro (ceftaroline fosamil),Adcirca (tadalafil), Tyvaso (treprostinil), Alvesco (ciclesonide),Patanase (olopatadine hydrochloride), Letairis (ambrisentan), Xyzal(levocetirizine dihydrochloride), Brovana (arformoterol tartrate),Tygacil (tigecycline), Ketek (telithromycin), Spiriva HandiHaler(tiotropium bromide), Aldurazyme (laronidase), Iressa (gefitinib),Xolair (omalizumab), Zemaira (alphal-proteinase inhibitor), Clarinex,Qvar (beclomethasone dipropionate), Remodulin (treprostinil), Xopenex,Avelox I.V. (moxifloxacin hydrochloride), DuoNeb (albuterol sulfate andipratropium bromide), Foradil Aerolizer (formoterol fumarate inhalationpowder), Invanz, NasalCrom Nasal Spray, Tavist (clemastine fumarate),Tracleer (bosentan), Ventolin HFA (albuterol sulfate inhalationaerosol), Biaxin XL (clarithromycin extended-release tablets), Cefazolinand Dextrose USP, Tri-Nasal Spray (triamcinolone acetonide spray),Accolate, Cafcit Injection, Proventil HFA Inhalation Aerosol, RhinocortAqua Nasal Spray, Tequin, Tikosyn Capsules, Allegra-D, Clemastinefumarate syrup, Curosurf, Dynabac, Infasurf, Priftin, Pulmozyme (dornasealfa), Sclerosol Intrapleural Aerosol, Singulair, Synagis, Ceftin(cefuroxime axetil), Cipro (ciprofloxacin HCl), Claritin RediTabs (10 mgloratadine rapidly-disintegrating tablet), Flonase Nasal Spray, FloventRotadisk, Metaprotereol Sulfate Inhalation Solution (5%), Nasacort AQ(triamcinolone acetonide) Nasal Spray, Omnicef, Raxar (grepafloxacin),Serevent, Tilade (nedocromil sodium), Tobi, Vanceril 84 mcg DoubleStrength (beclomethasone dipropionate, 84 mcg) Inhalation Aerosol, Zagam(sparfloxacin) tablets, Zyflo (Zileuton), Accolate, Allegra(fexofenadine hydrochloride), Astelin nasal spray, Atrovent (ipratropiumbromide), Augmentin (amoxicillin/clavulanate), Azmacort (triamcinoloneacetonide) Inhalation Aerosol, Breathe Right, Claritin Syrup(loratadine), Claritin-D 24 Hour Extended Release Tablets (10 mgloratadine, 240 mg pseudoephedrine sulfate), Covera-HS (verapamil),Nasacort AQ (triamcinolone acetonide) Nasal Spray, OcuHist, Pulmozyme(dornase alfa), RespiGam (Respiratory Syncitial Virus Immune GlobulinIntravenous), Tavist (clemastine fumarate), Tripedia (Diptheria andTetanus Toxoids and Acellular Pertussis Vaccine Absorbed), Vancenase AQ84 mcg Double Strength, Visipaque (iodixanol), Zosyn (sterilepiperacillin sodium/tazobactam sodium), Cedax (ceftibuten), and Zyrtec(cetirizine HCl). In one embodiment, the agent for pulmonary orrespiratory diseases is Arcapta, Daliresp, Dulera, Alvesco, Brovana,Spiriva HandiHaler, Xolair, Qvar, Xopenex, DuoNeb, Foradil Aerolizer,Accolate, Singulair, Flovent Rotadisk, Tilade, Vanceril, Zyflo, orAzmacort Inhalation Aerosol. In one embodiment, the agent for pulmonaryor respiratory diseases is Spiriva HandiHaler.

In some embodiments, a compound provided herein (e.g., compound 1), oran enantiomer or a mixture of enantiomers thereof, or a pharmaceuticallyacceptable salt, solvate, hydrate, co-crystal, clathrate, or polymorphthereof) is administered in combination with an agent for immunology orinfectious diseases. Examples of agents for immunology or infectiousdiseases include, but are not limited to, Horizant (gabapentinenacarbil), Qnasl (beclomethasone dipropionate) nasal aerosol, Rayos(prednisone) delayed-release tablets, Stribild (elvitegravir,cobicistat, emtricitabine, tenofovir disoproxil fumarate), TudorzaPressair (aclidinium bromide inhalation powder), Arcapta (indacaterolmaleate inhalation powder), Benlysta (belimumab), Complera(emtricitabine/rilpivirine/tenofovir disoproxil fumarate), Daliresp(roflumilast), Dificid (fidaxomicin), Edurant (rilpivirine), Firazyr(icatibant), Gralise (gabapentin), Incivek (telaprevir), Nulojix(belatacept), Victrelis (boceprevir), Cayston (aztreonam for inhalationsolution), Egrifta (tesamorelin for injection), Menveo (meningitisvaccine), Oravig (miconazole), Prevnar 13 (Pneumococcal 13-valentConjugate Vaccine), Teflaro (ceftaroline fosamil), Zortress(everolimus), Zymaxid (gatifloxacin ophthalmic solution), Bepreve(bepotastine besilate ophthalmic solution), Berinert (C1 EsteraseInhibitor (Human)), Besivance (besifloxacin ophthalmic suspension),Cervarix [Human Papillomavirus Bivalent (Types 16 and 18) Vaccine,Recombinant], Coartem (artemether/lumefantrine), Hiberix (Haemophilus bConjugate Vaccine; Tetanus Toxoid Conjugate), Ilaris (canakinumab),Ixiaro (Japanese Encephalitis Vaccine, Inactivated, Adsorbed), Kalbitor(ecallantide), Qutenza (capsaicin), Vibativ (telavancin), Zirgan(ganciclovir ophthalmic gel), Aptivus (tipranavir), Astepro (azelastinehydrochloride nasal spray), Cinryze (C1 Inhibitor (Human)), Intelence(etravirine), Moxatag (amoxicillin), Rotarix (Rotavirus Vaccine, Live,Oral), Tysabri (natalizumab), Viread (tenofovir disoproxil fumarate),Altabax (retapamulin), AzaSite (azithromycin), Doribax (doripenem),Extina (ketoconazole), Isentress (raltegravir), Selzentry (maraviroc),Veramyst (fluticasone furoate), Xyzal (levocetirizine dihydrochloride),Eraxis (anidulafungin), Gardasil (quadrivalent human papillomavirus(types 6, 11, 16, 18) recombinant vaccine), Noxafil (posaconazole),Prezista (darunavir), Rotateq (rotavirus vaccine, live oralpentavalent), Tyzeka (telbivudine), Veregen (kunecatechins), Aptivus(tipranavir), Baraclude (entecavir), Tygacil (tigecycline), Ketek(telithromycin), Tindamax, tinidazole, Xifaxan (rifaximin), Amevive(alefacept), FluMist (Influenza Virus Vaccine), Fuzeon (enfuvirtide),Lexiva (fosamprenavir calcium), Reyataz (atazanavir sulfate), Alinia(nitazoxanide), Clarinex, Daptacel, Fluzone Preservative-free, Hepsera(adefovir dipivoxil), Pediarix Vaccine, Pegasys (peginterferon alfa-2a),Restasis (cyclosporine ophthalmic emulsion), Sustiva, Vfend(voriconazole), Avelox I.V. (moxifloxacin hydrochloride), Cancidas,Peg-Intron (peginterferon alfa-2b), Rebetol (ribavirin), Spectracef,Twinrix, Valcyte (valganciclovir HCl), Viread (tenofovir disoproxilfumarate), Xigris (drotrecogin alfa [activated]), ABREVA (docosanol),Biaxin XL (clarithromycin extended-release tablets), Cefazolin andDextrose USP, Children's Motrin Cold, Evoxac, Kaletra Capsules and OralSolution, Lamisil (terbinafine hydrochloride) Solution (1%), Lotrisone(clotrimazole/betamethasone diproprionate) lotion, Malarone (atovaquone;proguanil hydrochloride) Tablet, Rapamune (sirolimus) Tablets, RidMousse, Tri-Nasal Spray (triamcinolone acetonide spray), Trivagizole 3(clotrimazole) Vaginal Cream, Trizivir (abacavir sulfate; lamivudine;zidovudine AZT) Tablet, Agenerase (amprenavir), Cleocin (clindamycinphosphate), Famvir (famciclovir), Norvir (ritonavir), Panretin Gel,Rapamune (sirolimus) oral solution, Relenza, Synercid I.V., Tamiflucapsule, Vistide (cidofovir), Allegra-D, CellCept, Clemastine fumaratesyrup, Cleocin (clindamycin phosphate), Dynabac, REBETRON™ CombinationTherapy, Simulect, Timentin, Viroptic, INFANRIX (Diphtheria and TetanusToxoids and Acellular Pertussis Vaccine Adsorbed), Acyclovir Capsules,Aldara (imiquimod), Aphthasol, Combivir, Condylox Gel 0.5% (pokofilox),Famvir (famciclovir), Flagyl ER, Flonase Nasal Spray, Fortovase,INFERGEN (interferon alfacon-1), Intron A (interferon alfa-2b,recombinant), Norvir (ritonavir), Rescriptor Tablets (delavirdinemesylate tablets), SPORANOX (itraconazole), Stromectol (ivermectin),Taxol, Trovan, VIRACEPT (nelfinavir mesylate), Zerit (stavudine),Albenza (albendazole), Apthasol (Amlexanox), Carrington patch, Confide,Crixivan (Indinavir sulfate), Gastrocrom Oral Concentrate (cromolynsodium), Havrix, Lamisil (terbinafine hydrochloride) Tablets, Leukine(sargramostim), Oral Cytovene, RespiGam (Respiratory Syncitial VirusImmune Globulin Intravenous), Videx (didanosine), Viramune (nevirapine),Vistide (cidofovir), Vitrasert Implant, Zithromax (azithromycin), Cedax(ceftibuten), Clarithromycin (Biaxin), Epivir (lamivudine), Intron A(Interferon alfa-2b, recombinant), Invirase (saquinavir), Valtrex(valacyclovir HCl), Western blot confirmatory device, Zerit (stavudine),and Zyrtec (cetirizine HCl).

In some embodiments, the second agent is an HDAC inhibitor, such as,e.g., belinostat, vorinostat, panobinostat, ACY-1215, or romidepsin.

In some embodiments, the second agent is an mTOR inhibitor, such as,e.g., everolimus (RAD 001).

In some embodiments, the second agent is a proteasome inhibitor, suchas, e.g., bortezomib or carfilzomib.

In some embodiments, the second agent is a PKC-β inhibitor, such as,e.g., Enzastaurin (LY317615).

In some embodiments, the second agent is a JAK/STAT inhibitor, such as,e.g., INCB16562 or AZD1480.

In some embodiments, the second agent is an anti-folate, such as, e.g.,pralatrexate.

In some embodiments, the second agent is a farnesyl transferaseinhibitor, such as, e.g., tipifarnib.

In some embodiments, the second agent is an antibody or a biologicagent, such as, e.g., alemtuzumab, rituximab, ofatumumab, or brentuximabvedotin (SGN-035). In one embodiment, the second agent is rituximab. Inone embodiment, the second agent is rituximab and the combinationtherapy is for treating, preventing, and/or managing iNHL, FL, splenicmarginal zone, nodal marginal zone, extranodal marginal zone, and/orSLL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination bendamustine and one additional active agent. Inone embodiment, the cancer or hematological malignancy is iNHL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination rituximab and one additional active agent. In oneembodiment, the cancer or hematological malignancy is iNHL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination bendamustine and rituximab. In one embodiment,the cancer or hematological malignancy is iNHL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination fludarabine, cyclophosphamide, and rituximab. Inone embodiment, the cancer or hematological malignancy is CLL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with an antibody or a biologic agent, such as,e.g., alemtuzumab, rituximab, ofatumumab, or brentuximab vedotin(SGN-035). In one embodiment, the second agent is rituximab. In oneembodiment, the second agent is rituximab and the combination therapy isfor treating, preventing, and/or managing iNHL, FL, splenic marginalzone, nodal marginal zone, extranodal marginal zone, and/or SLL.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with an antibody-drug conjugate, such as, e.g.,inotuzumab ozogamicin, or brentuximab vedotin.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with a cytotoxic agent, such as, e.g.,bendamustine, gemcitabine, oxaliplatin, cyclophosphamide, vincristine,vinblastine, anthracycline (e.g., daunorubicin or daunomycin,doxorubicin), actinomycin, dactinomycin, bleomycin, clofarabine,nelarabine, cladribine, asparaginase, methotrexate, or pralatrexate.

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with one or more other anti-cancer agents orchemotherapeutic agents, such as, e.g., fludarabine, ibrutinib,fostamatinib, lenalidomide, thalidomide, rituximab, cyclophosphamide,doxorubicin, vincristine, prednisone, or R-CHOP (Rituximab,Cyclophosphamide, Doxorubicin or Hydroxydaunomycin, Vincristine orOncovin, Prednisone).

In some embodiments, a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,is used in combination with an antibody for a cytokine (e.g., an IL-15antibody, an IL-21 antibody, an IL-4 antibody, an IL-7 antibody, an IL-2antibody, an IL-9 antibody). In some embodiments, the second agent is aJAK1 inhibitor, a JAK3 inhibitor, a pan-JAK inhibitor, a BTK inhibitor,an SYK inhibitor, or a PI3K-delta inhibitor. In some embodiments, thesecond agent is an antibody for a chemokine.

Without being limited to a particular theory, a targeted combinationtherapy described herein has reduced side effect and/or enhancedefficacy. For example, in one embodiment, provided herein is acombination therapy for treating CLL with a compound described herein(e.g., compound 1), or a pharmaceutically acceptable derivative (e.g.,salt or solvate) thereof, and a second active agent (e.g., IL-15antibodies, IL-21 antibodies, IL-4 antibodies, IL-7 antibodies, IL-2antibodies, IL-9 antibodies, JAK1 inhibitors, JAK3 inhibitors, pan-JAKinhibitors, BTK inhibitors, SYK inhibitors, and/or PI3K-deltainhibitors).

Further without being limited by a particular theory, it was found thata compound provided herein (e.g., compound 1) does not affect BTK or MEKpathway. Accordingly, in some embodiments, provided herein is a methodof treating or managing cancer or hematological malignancy comprisingadministering to a patient a therapeutically effective amount of acompound provided herein (e.g., compound 1), or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, in combinationwith a BTK inhibitor. In one embodiment, the BTK inhibitor is ibrutinib.In one embodiment, the BTK inhibitor is AVL-292. In one embodiment, thecancer or hematological malignancy is DLBCL. In another embodiment, thecancer or hematological malignancy is iNHL. In another embodiment, thecancer or hematological malignancy is CLL.

In other embodiments, provided herein is a method of treating ormanaging cancer or hematological malignancy comprising administering toa patient a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, in combination with a MEK inhibitor. Inone embodiment, the MEK inhibitor is trametinib/GSK1120212(N-(3-{3-Cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl}phenyl)acetamide),selumetinob(6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide),pimasertib/AS703026/MSC1935369((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide),XL-518/GDC-0973(1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(25)-piperidin-2-yl]azetidin-3-ol),refametinib/BAY869766/RDEA119(N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide),PD-0325901(N-[(2R)-2,3-Dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide),TAK733((R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione),MEK162/ARRY438162(5-[(4-Bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide),R05126766(3[[3-Fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one),WX-554, RO4987655/CH4987655(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazinan-2-yl)methyl)benzamide),or AZD8330(2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide).In one embodiment, the cancer or hematological malignancy is DLBCL. Inanother embodiment, the cancer or hematological malignancy is ALL. Inanother embodiment, the cancer or hematological malignancy is CTCL.

In other embodiments, provided herein is a method of treating ormanaging cancer or hematological malignancy comprising administering toa patient a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, in combination with an EZH2 inhibitor.In one embodiment, the EZH2 inhibitor is EPZ-6438, GSK-126, GSK-343,E11, or 3-deazaneplanocin A (DNNep). In one embodiment, the cancer orhematological malignancy is DLBCL. In another embodiment, the cancer orhematological malignancy is iNHL. In another embodiment, the cancer orhematological malignancy is ALL. In another embodiment, the cancer orhematological malignancy is CTCL.

In other embodiments, provided herein is a method of treating ormanaging cancer or hematological malignancy comprising administering toa patient a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, in combination with a bcl-2 inhibitor.In one embodiment, the BCL2 inhibitor is ABT-199(4-[4-[[2-(4-Chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide),ABT-737(4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide),ABT-263((R)-4-(4-((4′-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide),GX15-070 (obatoclax mesylate,(2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole;methanesulfonic acid))), or G3139 (Oblimersen). In one embodiment, thecancer or hematological malignancy is DLBCL. In another embodiment, thecancer or hematological malignancy is iNHL. In another embodiment, thecancer or hematological malignancy is CLL. In another embodiment, thecancer or hematological malignancy is ALL. In another embodiment, thecancer or hematological malignancy is CTCL.

In other embodiments, provided herein is a method of treating ormanaging iNHL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab. In one embodiment, the patient is anelderly patient. In another embodiment, iNHL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging iNHL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with bendamustine. In one embodiment, iNHL is relapsed orrefractory.

In other embodiments, provided herein is a method of treating ormanaging iNHL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab, and in further combination withbendamustine. In one embodiment, iNHL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging iNHL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with lenalidomide. In one embodiment, iNHL is relapsed orrefractory.

In other embodiments, provided herein is a method of treating ormanaging CLL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab. In one embodiment, the patient is anelderly patient. In another embodiment, CLL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging CLL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with bendamustine. In one embodiment, CLL is relapsed orrefractory.

In other embodiments, provided herein is a method of treating ormanaging CLL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab, and in further combination withbendamustine. In one embodiment, CLL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging CLL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with lenalidomide. In one embodiment, CLL is relapsed orrefractory.

In other embodiments, provided herein is a method of treating ormanaging DLBCL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab. In one embodiment, the patient is anelderly patient. In another embodiment, DLBCL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging DLBCL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with bendamustine. In one embodiment, DLBCL is relapsedor refractory.

In other embodiments, provided herein is a method of treating ormanaging DLBCL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with rituximab, and in further combination withbendamustine. In one embodiment, DLBCL is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging DLBCL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with R-GDP (rituximab, cyclophosphamide, vincristine andprednisone). In one embodiment, DLBCL is relapsed or refractory. Inanother embodiment, the treatment is done subsequent to treatment byR-CHOP.

In other embodiments, provided herein is a method of treating ormanaging DLBCL comprising administering to a patient a therapeuticallyeffective amount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with ibrutinib. In one embodiment, DLBCL is relapsed orrefractory.

In other embodiments, provided herein is a method of treating ormanaging T-cell lymphoma (PTCL or CTCL) comprising administering to apatient a therapeutically effective amount of a compound provided herein(e.g., compound 1), or a pharmaceutically acceptable derivative (e.g.,salt or solvate) thereof, in combination with rituximab. In oneembodiment, T-cell lymphoma is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging T-cell lymphoma (PTCL or CTCL) comprising administering to apatient a therapeutically effective amount of a compound provided herein(e.g., compound 1), or a pharmaceutically acceptable derivative (e.g.,salt or solvate) thereof, in combination with bendamustine. In oneembodiment, T-cell lymphoma is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging T-cell lymphoma (PTCL or CTCL) comprising administering to apatient a therapeutically effective amount of a compound provided herein(e.g., compound 1), or a pharmaceutically acceptable derivative (e.g.,salt or solvate) thereof, in combination with rituximab, and in furthercombination with bendamustine. In one embodiment, T-cell lymphoma isrelapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging T-cell lymphoma (PTCL or CTCL) comprising administering to apatient a therapeutically effective amount of a compound provided herein(e.g., compound 1), or a pharmaceutically acceptable derivative (e.g.,salt or solvate) thereof, in combination with romidepsin. In oneembodiment, T-cell lymphoma is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging mantle cell lymphoma comprising administering to a patient atherapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with rituximab. In one embodiment,mantle cell lymphoma is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging mantle cell lymphoma comprising administering to a patient atherapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with bendamustine. In one embodiment,mantle cell lymphoma is relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging mantle cell lymphoma comprising administering to a patient atherapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with rituximab, and in furthercombination with bendamustine. In one embodiment, mantle cell lymphomais relapsed or refractory.

In other embodiments, provided herein is a method of treating ormanaging mantle cell lymphoma comprising administering to a patient atherapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with ibrutinib. In one embodiment,mantle cell lymphoma is relapsed or refractory.

Further, without being limited by a particular theory, it was found thatcancer cells exhibit differential sensitivity profiles to doxorubicinand compounds provided herein. Thus, provided herein is a method oftreating or managing cancer or hematological malignancy comprisingadministering to a patient a therapeutically effective amount of acompound provided herein (e.g., compound 1), or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, in combinationwith a doxorubicin. In one embodiment, the cancer or hematologicalmalignancy is ALL.

In some embodiments, provided herein is a method of treating or managingcancer or hematological malignancy comprising administering to a patienta therapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with a AraC. In one embodiment, thecancer or hematological malignancy is AML.

In specific embodiments, compound 1 or a pharmaceutically acceptableform thereof, is used in combination with one or more second agent orsecond therapy provided herein.

In some embodiments, the second agent is an antibody-drug conjugate,such as, e.g., inotuzumab ozogamicin, or brentuximab vedotin.

In some embodiments, the second agent is a cytotoxic agent, such as,e.g., bendamustine, gemcitabine, oxaliplatin, cyclophosphamide,vincristine, vinblastine, anthracycline (e.g., daunorubicin ordaunomycin, doxorubicin), actinomycin, dactinomycin, bleomycin,clofarabine, nelarabine, cladribine, asparaginase, methotrexate, orpralatrexate.

In some embodiments, the second agent is one or more other anti-canceragents or chemotherapeutic agents, such as, e.g., fludarabine,ibrutinib, fostamatinib, lenalidomide, thalidomide, rituximab,cyclophosphamide, doxorubicin, vincristine, prednisone, or R-CHOP(Rituximab, Cyclophosphamide, Doxorubicin or Hydroxydaunomycin,Vincristine or Oncovin, Prednisone).

In some embodiments, the second agent is an antibody for a cytokine(e.g., an IL-15 antibody, an IL-21 antibody, an IL-4 antibody, an IL-7antibody, an IL-2 antibody, an IL-9 antibody). In some embodiments, thesecond agent is a JAK1 inhibitor, a JAK3 inhibitor, a pan-JAK inhibitor,a BTK inhibitor, an SYK inhibitor, or a PI3K-delta inhibitor. In someembodiments, the second agent is an antibody for a chemokine.

Without being limited to a particular theory, a targeted combinationtherapy described herein has reduced side effect and/or enhancedefficacy. For example, in one embodiment, provided herein is acombination therapy for treating CLL with a compound described herein(e.g., compound 1) and a second active agent (e.g., IL-15 antibodies,IL-21 antibodies, IL-4 antibodies, IL-7 antibodies, IL-2 antibodies,IL-9 antibodies, JAK1 inhibitors, JAK3 inhibitors, pan-JAK inhibitors,BTK inhibitors, SYK inhibitors, and/or PI3K-delta inhibitors).

Further without being limited by a particular theory, it was found thata compound provided herein (e.g., compound 1) does not affect BTK or MEKpathway. Accordingly, in some embodiments, provided herein is a methodof treating or managing cancer or hematological malignancy comprisingadministering to a patient a therapeutically effective amount of acompound provided herein (e.g., compound 1), or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, in combinationwith a BTK inhibitor. In one embodiment, the BTK inhibitor is ibrutinib.In one embodiment, the BTK inhibitor is AVL-292. In one embodiment, thecancer or hematological malignancy is DLBCL. In another embodiment, thecancer or hematological malignancy is CLL.

In other embodiments, provided herein is a method of treating ormanaging cancer or hematological malignancy comprising administering toa patient a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, in combination with a MEK inhibitor. Inone embodiment, the MEK inhibitor is trametinib, selumetinob,AS703026/MSC1935369, XL-518/GDC-0973, BAY869766/RDEA119, GSK1120212(trametinib), pimasertib, refametinib, PD-0325901, TAK733,MEK162/ARRY438162, R05126766, WX-554, R04987655/CH4987655 or AZD8330. Inone embodiment, the cancer or hematological malignancy is DLBCL. Inanother embodiment, the cancer or hematological malignancy is ALL. Inanother embodiment, the cancer or hematological malignancy is CTCL.

In other embodiments, provided herein is a method of treating ormanaging cancer or hematological malignancy comprising administering toa patient a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, in combination with a bcl-2 inhibitor.In one embodiment, the BCL2 inhibitor is ABT-199, ABT-737, ABT-263,GX15-070 (obatoclax mesylate) or G3139 (Genasense). In one embodiment,the cancer or hematological malignancy is DLBCL. In another embodiment,the cancer or hematological malignancy is ALL. In another embodiment,the cancer or hematological malignancy is CTCL.

Further, without being limited by a particular theory, it was found thatcancer cells exhibit differential sensitivity profiles to doxorubicinand compounds provided herein. Thus, provided herein is a method oftreating or managing cancer or hematological malignancy comprisingadministering to a patient a therapeutically effective amount of acompound provided herein (e.g., compound 1), or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, in combinationwith a doxorubicin. In one embodiment, the cancer or hematologicalmalignancy is ALL.

In some embodiments, provided herein is a method of treating or managingcancer or hematological malignancy comprising administering to a patienta therapeutically effective amount of a compound provided herein (e.g.,compound 1), or a pharmaceutically acceptable derivative (e.g., salt orsolvate) thereof, in combination with a AraC. In one embodiment, thecancer or hematological malignancy is AML.

In specific embodiments, compound 1 or a pharmaceutically acceptableform thereof, is used in combination with one or more second agent orsecond therapy provided herein.

In certain embodiments, provided herein are pharmaceutical compositioncomprising a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable formthereof, and a PI3K-delta inhibitor.

In one embodiment, the PI3K-delta inhibitor is a PI3K-delta selectiveinhibitor. In one embodiment, the PI3K-delta inhibitor is GS-1101(Cal-101), GSK-2269557, GS-9820, AMG319, or TGR-1202, or a mixturethereof. In one embodiment, the PI3K-delta inhibitor is of the formula:

or a pharmaceutically acceptable form thereof.

In one embodiment, the molar ratio of the compound, or apharmaceutically acceptable form thereof, to the PI3K-delta inhibitor isin the range of from about 10000:1 to about 1:10000. In one embodiment,the molar ratio of the compound, or a pharmaceutically acceptable formthereof, to the PI3K-delta inhibitor is in the range of from about 10:1to about 1:10. In one embodiment, the composition comprises thecompound, or a pharmaceutically acceptable form thereof, at an amount ofin the range of from about 0.01 mg to about 75 mg and the PI3K-deltainhibitor at an amount of in the range of from about 0.01 mg to about1100 mg. In one embodiment, the compound, or a pharmaceuticallyacceptable form thereof, and the PI3K-delta inhibitor are the onlytherapeutically active ingredients.

In one embodiment, the compound, or pharmaceutically acceptable formthereof, and the PI3K-delta inhibitor are in a single dosage form. Inone embodiment, the compound, or pharmaceutically acceptable formthereof, and the PI3K-delta inhibitor are in separate dosage forms. Inone embodiment, the composition further comprising a pharmaceuticallyacceptable excipient.

In one embodiment, the composition is synergistic in treating a cancer,inflammatory disease, or autoimmune disease.

In one embodiment, provided herein is a method of treating aPI3K-mediated disorder in a subject, comprising administering to thesubject a therapeutically effective amount of the composition.

In certain embodiments, provided herein are methods treating aPI3K-mediated disorder in a subject, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(e.g., compound 1), or a pharmaceutically acceptable form thereof, incombination with a PI3K-delta inhibitor.

In one embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered concurrently with the PI3K-delta inhibitor. Inone embodiment, the compound, or a pharmaceutically acceptable formthereof, is administered subsequent to the PI3K-delta inhibitor. In oneembodiment, the compound, or a pharmaceutically acceptable form thereof,is administered prior to the PI3K-delta inhibitor. In one embodiment,the compound, or a pharmaceutically acceptable form thereof, isadministered alone after discontinuing the administration of thePI3K-delta inhibitor.

In one embodiment, the PI3K-mediated disorder is a cancer, autoimmunedisease, or inflammatory disease. In one embodiment, the cancer is ofhematopoietic origin. In one embodiment, the cancer is a leukemia orlymphoma. In one embodiment, the leukemia or lymphoma is a B-celllymphoma, T-cell lymphoma, non-Hodgkin lymphoma, Hodgkin lymphoma, oranaplastic large cell lymphoma.

In one embodiment, the cancer is a solid tumor. In one embodiment, thecancer is selected from one or more of: a cancer of the pulmonarysystem, a brain cancer, a cancer of the gastrointestinal tract, a skincancer, a genitourinary cancer, a pancreatic cancer, a lung cancer, amedulloblastoma, a basal cell carcinoma, a glioma, a breast cancer, aprostate cancer, a testicular cancer, an esophageal cancer, ahepatocellular cancer, a gastric cancer, a gastrointestinal stromaltumor (GIST), a colon cancer, a colorectal cancer, an ovarian cancer, amelanoma, a neuroectodermal tumor, head and neck cancer, a sarcoma, asoft-tissue sarcoma, fibrosarcoma, myxosarcoma, liposarcoma, achondrosarcoma, an osteogenic sarcoma, a chordoma, an angiosarcoma, anendotheliosarcoma, a lymphangiosarcoma, a lymphangioendotheliosarcoma, asynovioma, a mesothelioma, a leiomyosarcoma, a cervical cancer, auterine cancer, an endometrial cancer, a carcinoma, a bladder carcinoma,an epithelial carcinoma, a squamous cell carcinoma, an adenocarcinoma, abronchogenic carcinoma, a renal cell carcinoma, a hepatoma, a bile ductcarcinoma, a neuroendocrine cancer, a carcinoid tumor, diffuse typegiant cell tumor, and glioblastoma.

In one embodiment, the PI3K-delta inhibitor is a PI3K-delta selectiveinhibitor. In one embodiment, the PI3K-delta inhibitor is of theformula:

or a pharmaceutically acceptable form thereof.

In one embodiment, the compound, or a pharmaceutically acceptable formthereof, and the PI3K-delta inhibitor are in a single dosage form. Inone embodiment, the compound, or a pharmaceutically acceptable formthereof, and the PI3K-delta inhibitor are in separate dosage forms.

In one embodiment, the concentration of the compound that is required toachieve 50% inhibition is at least 20%, 30%, 40%, or 50% lower when thecompound is administered in combination with the PI3K-delta inhibitorthan when the compound is administered individually. In one embodiment,the concentration of the PI3K-delta inhibitor that is required toachieve 50% inhibition is at least 20%, 30%, 40%, or 50% lower when thePI3K-delta inhibitor is administered in combination with the compoundthan when the PI3K-delta inhibitor is administered individually. In oneembodiment, the dose of the compound that is required to achieve 50%inhibition is at least 20%, 30%, 40%, or 50% lower when the compound isadministered in combination with the PI3K-delta inhibitor than when thecompound is administered individually. In one embodiment, the dose ofthe PI3K-delta inhibitor that is required to achieve 50% inhibition isat least 20%, 30%, 40%, or 50% lower when the PI3K-delta inhibitor isadministered in combination with the compound than when the PI3K-deltainhibitor is administered individually.

In one embodiment, the combination is synergistic as indicated by acombination index value that is less than 0.7, 0.5, or 0.1 for thecombination of the compound and the PI3K-delta inhibitor. In oneembodiment, the combination index value is assessed at 50% inhibition.In one embodiment, the combination index value is assessed at 50% growthinhibition. In one embodiment, the combination is synergistic asindicated by a Synergy Score that is greater than 1, 2, or 3 for thecombination of the Compound 1 and the PI3K-delta inhibitor. In oneembodiment, the combination is synergistic as indicated by a SynergyScore that is greater than 1, 2, or 3, for the combination of thecompound and the PI3K-delta inhibitor for inhibition or growthinhibition.

In one embodiment, the PI3K-mediated disorder is cancer, and theanti-cancer effect provided by the combination is at least 2 foldgreater, at least 3 fold greater, at least 5 fold greater, or at least10 fold greater than the anti-cancer effect provided by Compound 1, orpharmaceutically acceptable form thereof, alone. In one embodiment, thePI3K-mediated disorder is cancer, and the anti-cancer effect provided bythe combination is at least 2 fold greater, at least 3 fold greater, atleast 5 fold greater, or at least 10 fold greater than the anti-cancereffect provided by the PI3K-delta inhibitor alone.

In one embodiment, wherein one or more side effects associated withadministration of the compound, or a pharmaceutically acceptable formthereof, alone is reduced when the combination is administered at a dosethat achieves the same therapeutic effect. In one embodiment, one ormore side effects associated with administration of the PI3K-deltainhibitor alone is reduced when the combination is administered at adose that achieves the same therapeutic effect.

Combinations with Immune Modulators

While not wishing to be bound by theory, it is believed that tumorgrowth is influenced by at least two classes of immune cells in thetumor microenvironment: effector cells (including cytotoxic cells and M1macrophages) which have anti-tumor activity, and tumor associatedsuppressor cells (including M2 macrophages, MDSC, Tregs, and regulatorydendritic cells) which have pro-tumor activity because they inhibit theeffector cells or provide direct growth stimulation to the tumor cellsor tumor vasculature. An abundance of suppressor cells can lead to tumorimmune tolerance, and enhancement of tumor growth. A combination cancertherapy can be designed taking this mechanism into consideration.

For example, in embodiments, a PI3K-γ inhibitor as described herein (ora compound provided herein (e.g., compound 1) is administered incombination with a second therapeutic that blocks homeostaticdown-regulation of a T cell response in an effector T cell. This secondagent may be an immune checkpoint therapy as described below. As anotherexample, in embodiments, a PI3K-γ inhibitor as described herein isadministered in combination with a second therapeutic that reduces oreliminates suppressive cells in the tumor microenvironment, e.g., maydeplete MDSCs, TAMs or M2 macrophages, or any combination thereof. Thisagent could comprise, e.g., a CSF1R inhibitor, a CCL2 inhibitor, a CXCR4inhibitor, a MEK inhibitor, or an MTOR inhibitor, or any combinationthereof. In some embodiments, the second agent is an immunotherapy suchas a tumor vaccine, e.g., a tumor vaccine described herein. In someembodiments, the second agent is a cell therapy, e.g., a dendritic cellor a chimeric T cell, e.g., as described herein. In some embodiments,the second agent is an interleukin, e.g., IL7, IL12, IL15, or IL21.According to non-limiting theory, some interleukins exert an anti-cancereffect by stimulating the growth of immune cell populations.

In another embodiment, a compound provided herein (e.g., compound 1) isadministered in combination with a vaccine, e.g., a cancer vaccine,(e.g., a dendritic cell renal carcinoma (DC-RCC) vaccine). In certainembodiments, the combination of compound and the DC-RCC vaccine is usedto treat a cancer, e.g., a cancer as described herein (e.g., a renalcarcinoma, e.g., metastatic renal cell carcinoma (RCC) or clear cellrenal cell carcinoma (CCRCC)).

In some embodiments, a compound provided herein, or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, is administeredin combination with one or more immune checkpoint therapies. In someembodiments, provided herein is a method of treating a cancer in asubject, comprising administering to the subject a PI3K-gamma inhibitoror a compound as described herein (e.g., compound 1) in combination withone or more immune checkpoint therapies (e.g., PD-1 or PD-L1inhibitors). In some embodiments, provided herein is a method oftreating a solid cancer in a subject, comprising administering to thesubject Compound 1, or a pharmaceutically acceptable form thereof, incombination with one or more of PD-1 or PD-L1 inhibitors. In oneembodiment, the cancer is melanoma, bladder cancer, head and neckcancer, lung cancer (e.g., non-small cell lung cancer), or renal cellcarcinoma. In one embodiment, the cancer is melanoma. In one embodiment,the cancer is bladder cancer. In one embodiment, the cancer is lungcancer. In one embodiment, the cancer is non-small cell lung cancer. Inone embodiment, the cancer is renal cell carcinoma. In one embodiment,the cancer is head and neck cancer. In one embodiment, the cancer isbreast cancer. In one embodiment, the cancer is triple-negative breastcancer. In one embodiment, the cancer is colon cancer. In oneembodiment, the cancer is glioblastoma. In one embodiment, the cancer isovarian cancer.

In some embodiments, the subject is naive to immunotherapy treatment. Insome embodiments, the subject is naive to radiation therapy treatment.In some embodiments, the subject is naive to chemotherapy treatment.

In some embodiments, the subject has been pre-treated or previouslytreated with one or more immunotherapy treatments. In one embodiment,the subject is responsive to the pre-treatment or previous treatmentwith the immunotherapy. In one embodiment, the immunotherapy treatmentis a checkpoint treatment such as a PD-1 or PD-L1 inhibitor. In oneembodiment, the subject is a smoker.

In one embodiment, the cancer is melanoma, and the subject has beenpre-treated or previously treated with one or more immunotherapytreatments. In one embodiment, the subject has been pre-treated orpreviously treated with two or more immunotherapy treatments.

In one embodiment, the cancer is head and neck cancer, lung cancer(e.g., non-small cell lung cancer), renal cell carcinoma, or bladdercancer, and the subject has been pre-treated or previously treated withone immunotherapy treatment.

In one embodiment, the cancer is breast cancer (e.g., triple-negativebreast cancer), ovarian cancer, glioblastoma, or colon cancer, and thesubject is naive to immunotherapy treatment.

In some embodiments, the immune checkpoint therapy inhibits CTLA-4,PD-1, or PD-L1, or any combination thereof. The immune checkpointtherapy may be, e.g., a small molecule or an antibody. In someembodiments, the immune checkpoint therapy is an antibody that inhibitsprogrammed cell death 1 (also known as PD-1). In another embodiment, theimmune checkpoint therapy is nivolumab (also known as Opdivo). In someembodiments, the immune checkpoint therapy is anti-PD-L1 (programmedcell death ligand 1, also known as cluster of differentiation 274(CD274)), anti-PDL2, or anti-CTLA-4 (cytotoxic T-lymphocyte antigen 4,also known as cluster of differentiation (CD152)) antibody. Certainanti-PD-1, anti-PD-L1, and anti-CTLA-4 antibodies have activity inpreclinical and clinical tumor models. Cancer Res; 73(12) Jun. 15, 2013;Curran M A et al. PNAS 2010; 107:4275-4280; Topalian et al. N Engl J Med2012; 366:2443-2454; Wolchok et al., 2013. NEJM 369.

There are two main types of immune checkpoint therapies: an activator ofa costimulatory molecule, and an inhibitor of an immune checkpointmolecule.

When the immune checkpoint therapy is an activator of a costimulatorymolecule, it may be, e.g., chosen from an agonist (e.g., an agonisticantibody or antigen-binding fragment thereof, or a soluble fusion) ofOX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB(CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7,NKp80, CD160, B7-H3 or CD83 ligand. In certain embodiments, the immunecheckpoint therapy is an inhibitor of OX40 or anti-OX40 ab.

In the second situation, the immune checkpoint therapy is an inhibitorof an immune checkpoint molecule, for instance, an inhibitor of PD-1,PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4and/or TGFR beta. For instance, the inhibitor of an immune checkpointmolecule may inhibit PD-1, PD-L1, LAG-3, TIM-3 or CTLA4, or anycombination thereof.

Inhibition of an inhibitory molecule can be performed at the DNA, RNA orprotein level. For example, an inhibitory nucleic acid (e.g., a dsRNA,siRNA or shRNA), can be used to inhibit expression of an inhibitorymolecule. In other embodiments, the inhibitor of an inhibitory signalis, a polypeptide e.g., a soluble ligand (e.g., PD-1-Ig or CTLA-4 Ig),or an antibody or antigen-binding fragment thereof, that binds to theinhibitory molecule; e.g., an antibody or fragment thereof (alsoreferred to herein as “an antibody molecule”) that binds to PD-1, PD-L1,PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/orTGFR beta, or a combination thereof.

The antibody molecule may be, e.g., a full antibody or fragment thereof(e.g., a Fab, F(ab′)₂, Fv, or a single chain Fv fragment (scFv)). Theantibody molecule may be, e.g., in the form of a bispecific antibodymolecule. In one embodiment, the bispecific antibody molecule has afirst binding specificity to PD-1 or PD-L1 and a second bindingspecificity, e.g., a second binding specificity to TIM-3, LAG-3, orPD-L2. In certain embodiments, the antibody molecule is administered byinjection (e.g., subcutaneously or intravenously) at a dose of about 1to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to5 mg/kg, or about 3 mg/kg. The dosing schedule can vary from e.g., oncea week to once every 2, 3, or 4 weeks.

In certain embodiments, the immune checkpoint therapy is an inhibitor ofPD-1, e.g., human PD-1. In another embodiment, the immune checkpointtherapy is an inhibitor of PD-L1, e.g., human PD-L1. In one embodiment,the inhibitor of PD-1 or PD-L1 is an antibody molecule to PD-1 or PD-L1.The PD-1 or PD-L1 inhibitor can be administered alone, or in combinationwith other immune checkpoint therapies, e.g., in combination with aninhibitor of LAG-3, TIM-3 or CTLA4. In some embodiments, the inhibitorof PD-1 or PD-L1, e.g., the anti-PD-1 or PD-L1 antibody molecule, isadministered in combination with a LAG-3 inhibitor, e.g., an anti-LAG-3antibody molecule. In another embodiment, the inhibitor of PD-1 orPD-L1, e.g., the anti-PD-1 or PD-L1 antibody molecule, is administeredin combination with a TIM-3 inhibitor, e.g., an anti-TIM-3 antibodymolecule. In yet other embodiments, the inhibitor of PD-1 or PD-L1,e.g., the anti-PD-1 antibody molecule, is administered in combinationwith a LAG-3 inhibitor, e.g., an anti-LAG-3 antibody molecule, and aTIM-3 inhibitor, e.g., an anti-TIM-3 antibody molecule. Also providedherein are other combinations of immune checkpoint therapies with a PD-1inhibitor (e.g., one or more of PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA,TIGIT, LAIR1, CD160, 2B4 and/or TGFR). Any of the PI3K inhibitormolecules known in the art or disclosed herein can be used in theaforesaid combinations of inhibitors of checkpoint molecule.

In some embodiments, the anti-PD-1 antibody is Nivolumab. Alternativenames for Nivolumab include MDX-1106, MDX-1106-04, ONO-4538, orBMS-936558. In some embodiments, the anti-PD-1 antibody is Nivolumab(CAS Registry Number: 946414-94-4). Nivolumab is a fully human IgG4monoclonal antibody which specifically blocks PD1. Nivolumab (clone 5C4)and other human monoclonal antibodies that specifically bind to PD1 aredisclosed in U.S. Pat. No. 8,008,449 and WO2006/121168.

In other embodiments, the anti-PD-1 antibody is Pembrolizumab.Pembrolizumab (Trade name KEYTRUDA formerly Lambrolizumab, also known asMerck 3745, MK-3475 or SCH-900475) is a humanized IgG4 monoclonalantibody that binds to PD1. Pembrolizumab is disclosed, e.g., in Hamid,O. et al. (2013) New England Journal of Medicine 369 (2): 134-44,WO2009/114335, and U.S. Pat. No. 8,354,509.

In some embodiments, the anti-PD-1 antibody is Pidilizumab. Pidilizumab(CT-011; Cure Tech) is a humanized IgG1k monoclonal antibody that bindsto PD1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodiesare disclosed in WO2009/101611. Other anti-PD1 antibodies are disclosedin U.S. Pat. No. 8,609,089, US 2010028330, and/or US 20120114649. Otheranti-PD1 antibodies include AMP 514 (Amplimmune).

In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., animmunoadhesin comprising an extracellular or PD-1 binding portion ofPD-L1 or PD-L2 fused to a constant region (e.g., an Fc region of animmunoglobulin sequence)). In some embodiments, the PD-1 inhibitor isAMP-224. In some embodiments, a PI3K inhibitor, e.g., a PI3K-γ inhibitoras described herein (e.g., Compound 1), is administered together with animmunoadhesin (e.g., an immunoadhesin comprising an extracellular orPD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (e.g.,an Fc region of an immunoglobulin sequence)). In some embodiments, thecombination therapy is used in a method of treating a cancer, asdescribed herein.

In some embodiments, the PD-L1 inhibitor is anti-PD-L1 antibody. In someembodiments, the anti-PD-L1 inhibitor is chosen from YW243.55.570,MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105.

In one embodiment, the PD-L1 inhibitor is MDX-1105. MDX-1105, also knownas BMS-936559, is an anti-PD-L1 antibody described in WO2007/005874.

In one embodiment, the PD-L1 inhibitor is YW243.55.S70. The YW243.55.S70antibody is an anti-PD-L1 described in WO 2010/077634 (heavy and lightchain variable region sequences shown in SEQ ID Nos. 20 and 21,respectively).

In one embodiment, the PD-L1 inhibitor is MDPL3280A (Genentech/Roche).MDPL3280A is a human Fc optimized IgG1 monoclonal antibody that binds toPD-L1. MDPL3280A and other human monoclonal antibodies to PD-L1 aredisclosed in U.S. Pat. No. 7,943,743 and U.S. Publication No.:20120039906.

In other embodiments, the PD-L2 inhibitor is AMP-224. AMP-224 is a PD-L2Fc fusion soluble receptor that blocks the interaction between PD1 andB7-H1 (B7-DCIg; Amplimmune; e.g., disclosed in WO2010/027827 andWO2011/066342).

In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibodymolecule. In one embodiment, the LAG-3 inhibitor is BMS-986016.

In some embodiments, the anti-PD-L1 binding antagonist is chosen fromYW243.55.570, MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105. MDX-1105,also known as BMS-936559, is an anti-PD-L1 antibody described inWO2007/005874. Antibody YW243.55.570 (heavy and light chain variableregion sequences shown in SEQ ID Nos. 20 and 21, respectively) is ananti-PD-L1 described in WO 2010/077634.

In some embodiments, the anti-PD-1 antibody is Nivolumab. Alternativenames for Nivolumab include MDX-1106, MDX-1106-04, ONO-4538, orBMS-936558. In some embodiments, the anti-PD-1 antibody is Nivolumab(CAS Registry Number: 946414-94-4). Nivolumab (also referred to asBMS-936558 or MDX1106; Bristol-Myers Squibb) is a fully human IgG4monoclonal antibody which specifically blocks PD-1. Nivolumab (clone5C4) and other human monoclonal antibodies that specifically bind toPD-1 are disclosed in U.S. Pat. No. 8,008,449, EP2161336 andWO2006/121168.

In some embodiments, the anti-PD-1 antibody is Pembrolizumab.Pembrolizumab (also referred to as Lambrolizumab, MK-3475, MK03475,SCH-900475 or KEYTRUDA®; Merck) is a humanized IgG4 monoclonal antibodythat binds to PD-1. Pembrolizumab and other humanized anti-PD-1antibodies are disclosed in Hamid, O. et al. (2013) New England Journalof Medicine 369 (2): 134-44, U.S. Pat. No. 8,354,509 and WO2009/114335.

Pidilizumab (CT-011; Cure Tech) is a humanized IgGlk monoclonal antibodythat binds to PD1. Pidilizumab and other humanized anti-PD-1 monoclonalantibodies are disclosed in WO2009/101611.

Other anti-PD1 antibodies include AMP 514 (Amplimmune), among others,e.g., anti-PD1 antibodies disclosed in U.S. Pat. No. 8,609,089, US2010028330, and/or US 20120114649.

In some embodiments, the anti-PD-L1 antibody is MSB0010718C. MSB0010718C(also referred to as A09-246-2; Merck Serono) is a monoclonal antibodythat binds to PD-L1. Pembrolizumab and other humanized anti-PD-L1antibodies are disclosed in WO2013/079174.

MDPL3280A (Genentech/Roche) is a human Fc optimized IgG1 monoclonalantibody that binds to PD-L1. MDPL3280A and other human monoclonalantibodies to PD-L1 are disclosed in U.S. Pat. No.: 7,943,743 and U.SPublication No.: 20120039906. Other anti-PD-L1 binding agents includeYW243.55.S70 (heavy and light chain variable regions are shown in SEQ IDNOs 20 and 21 in WO2010/077634) and MDX-1105 (also referred to asBMS-936559, and, e.g., anti-PD-L1 binding agents disclosed inWO2007/005874).

AMP-224 (B7-DCIg; Amplimmune; e.g., disclosed in WO2010/027827 andWO2011/066342), is a PD-L2 Fc fusion soluble receptor that blocks theinteraction between PD1 and B7-H1.

In some embodiments, the anti-LAG-3 antibody is BMS-986016. BMS-986016(also referred to as BMS986016; Bristol-Myers Squibb) is a monoclonalantibody that binds to LAG-3. BMS-986016 and other humanized anti-LAG-3antibodies are disclosed in US 2011/0150892, WO2010/019570, andWO2014/008218.

In certain embodiments, the combination therapies disclosed hereininclude a modulator of a costimulatory molecule or an inhibitorymolecule, e.g., a co-inhibitory ligand or receptor.

In one embodiment, the costimulatory modulator, e.g., agonist, of acostimulatory molecule is chosen from an agonist (e.g., an agonisticantibody or antigen-binding fragment thereof, or soluble fusion) ofOX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB(CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7,NKp80, CD160, B7-H3 or CD83 ligand.

In another embodiment, the combination therapies disclosed hereininclude a costimulatory molecule, e.g., an agonist associated with apositive signal that includes a costimulatory domain of CD28, CD27, ICOSand GITR.

Exemplary GITR agonists include, e.g., GITR fusion proteins andanti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, aGITR fusion protein described in U.S. Pat. No. 6,111,090, EuropeanPatent No.: 090505B1, U.S. Pat. No. 8,586,023, PCT Publication Nos.: WO2010/003118 and 2011/090754, or an anti-GITR antibody described, e.g.,in U.S. Pat. No. 7,025,962, European Patent No.: 1947183B1, U.S. Pat.No. 7,812,135, U.S. Pat. No. 8,388,967, U.S. Pat. No. 8,591,886,European Patent No.: EP 1866339, PCT Publication No.: WO 2011/028683,PCT Publication No.: WO 2013/039954, PCT Publication No.: WO2005/007190,PCT Publication No.: WO 2007/133822, PCT Publication No.: WO2005/055808,PCT Publication No.: WO 99/40196, PCT Publication No.: WO 2001/03720,PCT Publication No.: WO99/20758, PCT Publication No.: WO2006/083289, PCTPublication No.: WO 2005/115451, U.S. Pat. No. 7,618,632, and PCTPublication No.: WO 2011/051726.

In one embodiment, the inhibitor is a soluble ligand (e.g., aCTLA-4-Ig), or an antibody or antibody fragment that binds to PD-L1,PD-L2 or CTLA4. For example, a compound disclosed herein, e.g., Compound1, can be administered in combination with an anti-CTLA-4 antibody,e.g., ipilimumab, for example, to treat a cancer (e.g., a cancer chosenfrom: a melanoma, e.g., a metastatic melanoma; a lung cancer, e.g., anon-small cell lung carcinoma; or a prostate cancer). Exemplaryanti-CTLA4 antibodies include Tremelimumab (IgG2 monoclonal antibodyavailable from Pfizer, formerly known as ticilimumab, CP-675,206); andIpilimumab (CTLA-4 antibody, also known as MDX-010, Yervoy, CAS No.477202-00-9). In some embodiments, a compound provided herein isadministered in combination with an anti-PD-L1 inhibitor (e.g.,nivolumab) and a CTLA-4 antibody (e.g., ipilimumab). In someembodiments, a compound provided herein is administered in combinationwith nivolumab and ipilimumab.

In some embodiments, a compound provided herein, or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, is administeredin combination with an anti-PD-L1 or anti-CTLA-4 antibody. In someembodiments, a compound provided herein, or a pharmaceuticallyacceptable derivative (e.g., salt or solvate) thereof, is administeredin combination with an anti-PD-L1 antibody. In another embodiment, acompound provided herein, or a pharmaceutically acceptable derivative(e.g., salt or solvate) thereof, is administered in combination withanti-CTLA-4 antibody. In some embodiments, the anti-PD-L1 antibody isselected from BMS-936559, MPDL3280A, and MDX-1105. In some embodiments,the anti-CTLA-4 antibody is selected from ipilimumab and tremelimumab.

In some embodiments, provided herein is a method of treating breastcancer, colon cancer, pancreatic cancer, melanoma, glioblastoma, or lungcancer comprising administering to a patient a therapeutically effectiveamount of a compound provided herein (e.g., compound 1), or apharmaceutically acceptable derivative (e.g., salt or solvate) thereof,in combination with an anti-PD-L1 or an anti-CTLA-4 antibody. In anotherembodiment, the cancer is chosen form a carcinoma (e.g., advanced ormetastatic carcinoma), melanoma or a lung carcinoma, e.g., a non-smallcell lung carcinoma. In one embodiment, the cancer is a lung cancer,e.g., a non-small cell lung cancer. In one embodiment, the cancer is amelanoma, e.g., an advanced melanoma. In one embodiment, the cancer isan advanced or unresectable melanoma that does not respond to othertherapies. In other embodiments, the cancer is a melanoma with a BRAFmutation (e.g., a BRAF V600E mutation). In another embodiment, thecancer is a hepatocarcinoma, e.g., an advanced hepatocarcinoma, with orwithout a viral infection, e.g., a chronic viral hepatitis. In anotherembodiment, the cancer is a prostate cancer, e.g., an advanced prostatecancer. In yet another embodiment, the cancer is a myeloma, e.g.,multiple myeloma. In yet another embodiment, the cancer is a renalcancer, e.g., a renal cell carcinoma (RCC) (e.g., a metastatic RCC orclear cell renal cell carcinoma (CCRCC)).

For example, a compound provided herein (e.g., compound 1) can beadministered in combination with an anti-CTLA-4 antibody, e.g.,ipilimumab, for example, to treat a cancer (e.g., a cancer chosen from:a melanoma, e.g., a metastatic melanoma; a lung cancer, e.g., anon-small cell lung carcinoma; or a prostate cancer). In one embodiment,a compound provided herein (e.g., compound 1) is administered aftertreatment with an anti-CTLA4 antibody (e.g., ipilimumab) with or withouta BRAF inhibitor (e.g., vemurafenib or dabrafenib).

In some embodiments, the immune checkpoint therapy is a costimulatoryligand. In some embodiments, the costimulatory ligand is OX40L, 41BBL,CD153, ICOSL, or CD40L.

In some embodiments, the immune checkpoint therapy is a MCSF/CSF-1Rinhibitor. An anti-CSF-1R can deplete TAMs, resulting in tumor growthinhibition. Cancer Cell 25, 1-14, Jun. 16, 2014. In some embodiments,the CSF-1R inhibitor is BLZ945, GW2850, R05509554, or PLX3397. In someembodiments, the CSF-1R inhibitor is BLZ945 or GW2850. In someembodiments, the CSF-1R inhibitor is PLX3397.

In some embodiments, the immune checkpoint therapy is animmunostimulant. In some embodiments, the immunostimulant is GMCSF, TLRligands, 41BBL, or ICOSL.

In some embodiments, the immune checkpoint therapy is a CXCR4/CXCL12inhibitor. In some embodiments, the CXCR4/CXCL12 inhibitor is AMD3100,AMD11070, AMD12118, AMD11814, or AMD13073. In some embodiments, theCXCR4/CXCL12 inhibitor is AMD3100.

In some embodiments, the immunotherapy is a CCL2 and/or CCR2 antagonist.In some embodiments, the antagonist of CCL2 and/or CCR2 is an anti-CCL2or CCR2 antibody. CCL2 is a chemokine and CCR2 is a chemokine receptor.CCL2 and CCR2, according to non-limiting theory, play a role in MDSCmigration.

In some embodiments, a PI3K-γ inhibitor disclosed herein, e.g., Compound1, is administered in combination with a BTK inhibitor. In oneembodiment, the BTK inhibitor is BTK inhibitors such as ibrutinib,AVL-292, Dasatinib, LFM-AI3, ONO-WG-307, and GDC-0834.

In some embodiments, a PI3K-γ inhibitor disclosed herein, e.g., Compound1, is administered in combination with an IDO (indoleamine2,3-dioxygenase) inhibitor or an TDO (tryptophan 2,3-dioxygenase)inhibitor. In one embodiment, the IDO inhibitor is indoximod, NLG919,INCB024360, F001287, norharmane, rosmarinic acid, oralpha-methyl-tryptophan Although IDO inhibitors act within the TME, theydo not specifically target MDSCs. The overexpression of IDO by dendriticcells creates an immunosuppressive tumor microenvironment.

In some embodiments, a PI3K-γ inhibitor disclosed herein, e.g., Compound1, is administered in combination with an inhibitor of one or moremembers of TAM family, a receptor tyrosine kinase (RTK) subfamilycomprising Tyro-3 (also called Sky), Ax1 and Mer. In one embodiment, theTAM inhibitor is BGB324 (R428), S49076, TP0903, CEP-40783, ONO-9330547,bosutinib (SKI606, PF5208763), cabozantinib (XL184), sunitinib(SU11248), foretinib (XL880, GSK1363089), MGCD265, BMS777607 (ASLAN002),LY2801653, SGI7079, amuvatinib (SGI-0470-02, MP470), SNS314,PF-02341066, diaminopyrimidine, spiroindoline, UNC569, UNC1062, UNC1666,UNC2025, or LDC1267. Additional TAM inhibitors include those describedin Mollard et al., Med. Chem. Lett. 2011, 2, 907-912 and Feneyrolles etal., Mol. Cancer Ther. 13(9), Published OnlineFirst Aug. 19, 2014, theentireties of which are incorporated by reference herein.

In some embodiment, a PI3K-γ inhibitor disclosed herein, e.g., Compound1, is administered to a subject concurrent or prior to theadministration of immune checkpoint therapy. In some embodiment, animmunostimulant is administered to a subject concurrent or prior to theadministration of immune checkpoint therapy. In some embodiment,chemotherapy (e.g., carboplatin, oxaliplatin, or radiation) isadministered to a subject concurrent or prior to the administration ofimmune checkpoint therapy.

In some embodiments, a PI3K-γ inhibitor disclosed herein, e.g., Compound1, is administered in combination with an ARG1 inhibitor. While notwishing to be bound by theory, it has been reported that tumorassociated myeloid cells establish an immunosuppressive microenvironmentin tumors through the expression of Arginase-1, which depletes the tumormicroenvironment of arginine, thereby the death or inhibition ofanti-tumor immune cells. Schmid et al., Proceedings: AACR 103rd AnnualMeeting 2012, Cancer Research: Apr. 15, 2012; Volume 72, Issue 8,Supplement 1. It has been reported that suppression of PI3K-gamma orArginase-1 expression blocked myeloid cell induced death of T cells invitro. Id. According to the non-limiting theory, PI3K-gamma inhibitionblocks Arginase-1 expression, thereby increasing the number of CD8+Tcells in tumors, stimulating T cell-mediated cytotoxicity of tumorcells, and suppressing growth and metastasis of tumors. Combinationtherapies can be designed in accordance with this mechanism.

For instance, in some embodiments, a PI3K-γ inhibitor disclosed herein,e.g., Compound 1, is administered in combination with an ARG1 inhibitor.The ARG1 inhibitor may be, e.g., an inhibitory nucleic acid such as asiRNA, an inhibitory anti-ARG-1 antibody, or an analog of arginine.Other exemplary inhibitors of ARG1 include N-hydroxy-guanidinium orN-hydroxy-nor-1-arginine, and boronic acid derivatives, such as,2(S)-amino-6-boronohexanoic acid, and S-(2-boronoethyl)-1-cysteine,α-α-disubstituted amino acid based arginase inhibitors [such as(R)-2-amino-6-borono-2-(2-(piperidin-1-yl)ethyl)hexanoic acid], andpiceatannol-3′-O-β-d-glucopyranoside (PG). Steppan et al., “Developmentof novel arginase inhibitors for therapy of endothelial dysfunction.”,Front Immunol. 2013 Sep. 17; 4:278. doi: 10.3389/fimmu.2013.00278.

The PI3K-γ inhibitors disclosed herein can have minimal effects onT-cell activation when compared to the suppressive effect of a PI3K δinhibitor on T-cell activation. Lewis lung carcinoma tumor growth can bereduced in PI3K-γ knockout mice and can have decreased tumor associatedsuppressive myeloid cell infiltrates. Tumor associated suppressivemyeloid cells can include e.g., myeloid derived suppressor cells (MDSCs)and tumor associated macrophages (TAMs). PI3K-γ knockout mice have TAMswhere the M2 phenotype is lost. M2 cells are immunosuppressive andsupport tumor growth. PI3K inhibitors provided herein can block M2phenotype (e.g., in an in vitro differentiation system), and thus canslow tumor growth.

For example, the effect of PI3K-γ inhibitors and PI3K δ inhibitors on Tcell activation as measured by inhibition of IFN-γ in response to ConAhas shown that PI3K-δ is plays a role in mediating T cell activation,while PI3K-γ has minimal effects on T-cell activation. The IC₅₀ for aPI3K-δ inhibitor in this assay is 3 nM, and the IC₅₀ for a PI3K-γinhibitor is 2500 nM. Administration of PI3K-γ inhibitors can lead toimpaired. T-cell migration but may have reduced effects on T-cellproliferation or activation.

In some embodiments, the PI3K-γ inhibitors disclosed herein can havepotent effects on tumor associated suppressive myeloid cells withoutinhibiting the effector T-cell. The PI3K-γ inhibitors disclosed hereincan have potent effects on tumor associated suppressive myeloid cellswithout blocking anti-tumor T-cell effects and thus can increase T cellactivity. In one embodiment, this effect can be enhanced byadministering CTLA4 antagonists and/or PD-1 and PDL1 antagonists. ThePI3K-γ inhibitors disclosed herein can increase T cell activation andproliferation. In some embodiments, provided herein is a method ofblocking tumor associated suppressive myeloid cells without inhibitingthe effects on anti-tumor T-cells comprising administering an effectiveamount of a PI3K-γ inhibitor disclosed herein or a pharmaceuticallyacceptable salt thereof to a subject. In some embodiments, providedherein is a method of blocking tumor associated suppressive myeloidcells without inhibiting the effects on anti-tumor T-cells comprisingadministering an effective amount of a compound disclosed herein or apharmaceutically acceptable salt thereof to a subject. In someembodiments, the subject has lung cancer, breast cancer, glioblastoma,or lymphoma (e.g., non-Hodgkin's lymphoma).

Further provided herein are methods of modulating kinase activity bycontacting a kinase with an amount of a compound provided hereinsufficient to modulate the activity of the kinase. Modulate can beinhibiting or activating kinase activity. In some embodiments, providedherein are methods of inhibiting kinase activity by contacting a kinasewith an amount of a compound provided herein sufficient to inhibit theactivity of the kinase. In some embodiments, provided herein are methodsof inhibiting kinase activity in a solution by contacting said solutionwith an amount of a compound provided herein sufficient to inhibit theactivity of the kinase in said solution. In some embodiments, providedherein are methods of inhibiting kinase activity in a cell by contactingsaid cell with an amount of a compound provided herein sufficient toinhibit the activity of the kinase in said cell. In some embodiments,provided herein are methods of inhibiting kinase activity in a tissue bycontacting said tissue with an amount of a compound provided hereinsufficient to inhibit the activity of the kinase in said tissue. In someembodiments, provided herein are methods of inhibiting kinase activityin an organism by contacting said organism with an amount of a compoundprovided herein sufficient to inhibit the activity of the kinase in saidorganism. In some embodiments, provided herein are methods of inhibitingkinase activity in an animal by contacting said animal with an amount ofa compound provided herein sufficient to inhibit the activity of thekinase in said animal. In some embodiments, provided herein are methodsof inhibiting kinase activity in a mammal by contacting said mammal withan amount of a compound provided herein sufficient to inhibit theactivity of the kinase in said mammal. In some embodiments, providedherein are methods of inhibiting kinase activity in a human bycontacting said human with an amount of a compound provided hereinsufficient to inhibit the activity of the kinase in said human. In someembodiments, the % of kinase activity after contacting a kinase with acompound provided herein is less than 1, 5, 10, 20, 30, 40, 50, 60, 70,80 90, 95, or 99% of the kinase activity in the absence of saidcontacting step.

In certain embodiments, provided herein are pharmaceutical compositioncomprising a therapeutically effective amount of a compound providedherein (e.g., compound 1), or a pharmaceutically acceptable formthereof, and an immunomodulator.

In one embodiment, the immunomodulator is an inhibitor of PD-1, PD-L1,LD-L2, CTLA-4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4,TGFR-beta, or IDO/TDO, or a combination thereof. In one embodiment, theimmunomodulator is an inhibitor of PD-L1. In one embodiment, theimmunomodulator is an antibody or fragment thereof, an inhibitorynucleic acid, a soluble ligand, or a fusion of a PD-1 ligand with a Fcregion of an immunoglobulin. In one embodiment, the immunomodulator is acostimulatory ligand, a MCSF/CSF-1R inhibitor, an immunostimulant, aCXCR4/CXCL12 inhibitor, a CCL2 inhibitor, or a CCR2 inhibitor. In oneembodiment, the immunomodulator is cyclophosphamide, docetaxel,paclitaxel, 5-FU, or temozolomide.

In one embodiment, provided herein is a method of treating aPI3K-mediated disorder in a subject, comprising administering to thesubject a therapeutically effective amount of the composition.

Combination Therapy for Pulmonary and Respiratory Diseases

In some embodiments, the compound provided herein is administered incombination with one or more other therapies. Such therapies includetherapeutic agents as well as other medical interventions, behavioraltherapies (e.g., avoidance of sunlight), and the like.

By “in combination with,” it is not intended to imply that the othertherapy and the compound provided herein must be administered at thesame time and/or formulated for delivery together, although thesemethods of delivery are within the scope of this disclosure. Thecompound provided herein can be administered concurrently with, prior to(e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 12 weeks, or16 weeks before), or subsequent to (e.g., 5 minutes, 15 minutes, 30minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, 6 weeks, 8 weeks, 12 weeks, or 16 weeks after), one or moreother therapies (e.g., one or more other additional agents). In general,each therapeutic agent will be administered at a dose and/or on a timeschedule determined for that particular agent. The other therapeuticagent can be administered with the compound provided herein in a singlecomposition or separately in a different composition. Triple therapy isalso contemplated herein.

In general, it is expected that additional therapeutic agents employedin combination be utilized at levels that do not exceed the levels atwhich they are utilized individually. In some embodiments, the levelsutilized in combination will be lower than those utilized individually.

In some embodiments, the compound provided herein is a first linetreatment for a pulmonary or respiratory disease, i.e., it is used in asubject who has not been previously administered another drug intendedto treat a pulmonary or respiratory disease, or one or more symptoms ofthe disease.

In some embodiments, the compound provided herein is a second linetreatment for a pulmonary or respiratory disease, i.e., it is used in asubject who has been previously administered another drug intended totreat a pulmonary or respiratory disease, or one or more symptoms of thedisease.

In some embodiments, the compound provided herein is a third or fourthline treatment for a pulmonary or respiratory disease, i.e., it is usedin a subject who has been previously administered two or three otherdrugs intended to treat a pulmonary or respiratory disease, or one ormore symptoms of the disease.

In embodiments where two agents are administered, the agents can beadministered in any order. For example, the two agents can beadministered concurrently (i.e., essentially at the same time, or withinthe same treatment) or sequentially (i.e., one immediately following theother, or alternatively, with a gap in between administration of thetwo). In some embodiments, the compound provided herein is administeredsequentially (i.e., after the first therapeutic).

In some embodiments, the compound provided herein and the second agentare administered as separate compositions, e.g., pharmaceuticalcompositions. In some embodiments, the compound provided herein and theagent are administered separately, but via the same route (e.g., both byinhalation). In some embodiments, the compound provided herein and theagent are administered in the same composition, e.g., pharmaceuticalcomposition.

In some embodiments, the compound provided herein (e.g., PI3K-δinhibitor or PI3K-γ inhibitor) is administered in combination with anagent that inhibits IgE production or activity. In some embodiments, thecompound provided herein (e.g., PI3K-δ inhibitor or PI3K-γ inhibitor) isadministered in combination with an inhibitor of mTOR. Agents thatinhibit IgE production are known in the art and they include but are notlimited to one or more of TEI-9874,2-(4-(6-cyclohexyloxy-2-naphtyloxy)phenylacetamide)benzoic acid,rapamycin, rapamycin analogs (i.e., rapalogs), TORC1 inhibitors, TORC2inhibitors, and any other compounds that inhibit mTORC1 and mTORC2.Agents that inhibit IgE activity include, for example, anti-IgEantibodies such as for example Omalizumab and TNX-901.

In certain embodiments, wherein inflammation (e.g., COPD, asthma) istreated, prevented and/or managed, a compound provided herein can becombined with, for example: PI3K inhibitors such as RP-6530, TG 100-115,RV1729, GS-1101, XL 499, GDC-0941, and AMG-319; BTK inhibitors such asibrutinib and AVL-292; JAK inhibitors such as tofacitinib and GLPG0636;SYK inhibitors such as fostamatinib.

In some embodiments, a compound provided herein can be combined withother agents that act to relieve the symptoms of inflammatoryconditions, such as COPD, asthma, and the other diseases describedherein. These agents include, but are not limited to, non-steroidalanti-inflammatory drugs (NSAIDs), e.g., acetylsalicylic acid; ibuprofen;naproxen; indomethacin; nabumetone; and tolmetin. In some embodiments,corticosteroids are used to reduce inflammation and suppress activity ofthe immune system.

In some embodiments, a compound provided herein (e.g., Compound 1), oran enantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof, isadministered in combination with an agent for pulmonary or respiratorydiseases. Examples of agents for pulmonary or respiratory diseasesinclude, but are not limited to, Abraxane (paclitaxel protein-boundparticles for injectable suspension), Adempas (riociguat), Anoro Ellipta(umeclidinium and vilanterol inhalation powder), Breo Ellipta(fluticasone furoate and vilanterol inhalation powder), Opsumit(macitentan), Qnasl (beclomethasone dipropionate) nasal aerosol, Sirturo(bedaquiline), Dymista (azelastine hydrochloride and fluticasonepropionate), Kalydeco (ivacaftor), Qnasl (beclomethasone dipropionate)nasal aerosol, Rayos (prednisone) delayed-release tablets, Surfaxin(lucinactant), Tudorza Pressair (aclidinium bromide inhalation powder),Arcapta (indacaterol maleate inhalation powder), Daliresp (roflumilast),Xalkori (crizotinib), Cayston (aztreonam for inhalation solution),Dulera (mometasone furoate+formoterol fumarate dihydrate), Teflaro(ceftaroline fosamil), Adcirca (tadalafil), Tyvaso (treprostinil),Alvesco (ciclesonide), Patanase (olopatadine hydrochloride), Letairis(ambrisentan), Xyzal (levocetirizine dihydrochloride), Brovana(arformoterol tartrate), Tygacil (tigecycline), Ketek (telithromycin),Spiriva HandiHaler (tiotropium bromide), Aldurazyme (laronidase), Iressa(gefitinib), Xolair (omalizumab), Zemaira (alphal-proteinase inhibitor),Clarinex, Qvar (beclomethasone dipropionate), Remodulin (treprostinil),Xopenex (levalbuterol), Avelox I.V. (moxifloxacin hydrochloride), DuoNeb(albuterol sulfate and ipratropium bromide), Foradil Aerolizer(formoterol fumarate inhalation powder), Invanz, NasalCrom Nasal Spray,Tavist (clemastine fumarate), Tracleer (bosentan), Ventolin HFA(albuterol sulfate inhalation aerosol), Biaxin XL (clarithromycinextended-release tablets), Cefazolin and Dextrose USP, Tri-Nasal Spray(triamcinolone acetonide spray), Accolate (zafirlukast), CafcitInjection, Proventil HFA Inhalation Aerosol, Rhinocort Aqua Nasal Spray,Tequin, Tikosyn Capsules, Allegra-D, Clemastine fumarate syrup,Curosurf, Dynabac, Infasurf, Priftin, Pulmozyme (dornase alfa),Sclerosol Intrapleural Aerosol, Singulair (montelukast sodium), Synagis,Ceftin (cefuroxime axetil), Cipro (ciprofloxacin HCl), Claritin RediTabs(10 mg loratadine rapidly-disintegrating tablet), Flonase Nasal Spray,Flovent Rotadisk, Metaprotereol Sulfate Inhalation Solution (5%),Nasacort AQ (triamcinolone acetonide) Nasal Spray, Omnicef, Raxar(grepafloxacin), Serevent, Tilade (nedocromil sodium), Tobi, Vanceril 84mcg Double Strength (beclomethasone dipropionate, 84 mcg) InhalationAerosol, Zagam (sparfloxacin) tablets, Zyflo (Zileuton), Allegra(fexofenadine hydrochloride), Astelin nasal spray, Atrovent (ipratropiumbromide), Augmentin (amoxicillin/clavulanate), Azmacort (triamcinoloneacetonide) Inhalation Aerosol, Breathe Right, Claritin Syrup(loratadine), Claritin-D 24 Hour Extended Release Tablets (10 mgloratadine, 240 mg pseudoephedrine sulfate), Covera-HS (verapamil),OcuHist, RespiGam (Respiratory Syncitial Virus Immune GlobulinIntravenous), Tripedia (Diptheria and Tetanus Toxoids and AcellularPertussis Vaccine Absorbed), Vancenase AQ 84 mcg Double Strength,Visipaque (iodixanol), Zosyn (sterile piperacillin sodium/tazobactamsodium), Cedax (ceftibuten), and Zyrtec (cetirizine HCl). In oneembodiment, the agent for pulmonary or respiratory diseases is Arcapta(indacaterol maleate inhalation powder), Daliresp (roflumilast), Dulera(mometasone furoate+formoterol fumarate dihydrate), Alvesco(ciclesonide), Brovana (arformoterol tartrate), Spiriva HandiHaler(tiotropium bromide), Xolair (omalizumab), Qvar (beclomethasonedipropionate), Xopenex (levalbuterol), DuoNeb (albuterol sulfate andipratropium bromide), Foradil Aerolizer (formoterol fumarate inhalationpowder), Accolate (zafirlukast), Singulair (montelukast sodium), FloventRotadisk (fluticasone propionate inhalation powder), Tilade (nedocromilsodium), Vanceril (beclomethasone dipropionate, 84 mcg), Zyflo(Zileuton), and Azmacort (triamcinolone acetonide) Inhalation Aerosol.In one embodiment, the agent for pulmonary or respiratory diseases isSpiriva HandiHaler (tiotropium bromide).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, acetylcysteine (mucomyst) selected from TudorzaPressair (aclidinium bromide), Atrovent (ipratropium), and Spiriva(tiotropium).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, beta2 agonists selected from short-acting beta2agonists and long acting beta2 agonists. Short acting beta2 agonistsinclude, but are not limited to, Proventil (albuterol), Tomalate(bitolterol), Xopenex (levalbuterol), Maxair (pirbuterol), and Alupent(metaproterenol). Long acting beta2 agonists include, but are notlimited to, Brovana (arformoterol tartrate), Foradil (formoterol),Arcapta Neohaler (indacaterol maleate), and Serevent (salmeterol).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, combination of two agents. In one embodiment, thecombination is administered through inhalation. The combination of twoagents includes, but is not limited to a beta2 agonist and ananticholinergi selected from Combivent (albuterol and ipratropium) andAnoro Ellipta (umeclidinium and vilanterol inhalation powder). Thecombination of two agents include, but are not limited to a beta2agonist and a corticosteroid selected from Advair (fluticasone andsalmeterol), Breo Ellipta (fluticasone furoate and vilanterol inhalationpowder), Dulera (mometasone furoate and formoterol fumarate), andSymbicort (budesonide and formoterol).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, corticosteroids selected from Vanceril Beclovent(beclomethasone), Pulmicort (budesonide), Alvesco (ciclesonide), Aerobid(flunisolide), Flovent (fluticasone), Asmanex (mometasone furoate), andAzmacort (triamcinolone).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, leukotriene inhibitors selected from Singulair(montelukast), Accolate (zafirlukast), and Zyflo (zileuton).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, mast cell stablizers selected from Intal (cromolynsodium) and Tilade (nedocromil).

Examples of agents for pulmonary or respiratory diseases include, butare not limited to, phosphodiesterase 4 (PDE4) inhibitors selected fromDaliresp (roflumilast).

In some embodiments, a compound provided herein (e.g., Compound 1), oran enantiomer, a mixture of enantiomers, or a mixture of two or morediastereomers thereof, or a pharmaceutically acceptable form thereof, isadministered in combination with an agent for immunology or infectiousdiseases. Examples of agents for immunology or infectious diseasesinclude, but are not limited to, Kineret (anakinra), Lovenox (enoxaparinsodium) Injection, Makena (hydroxyprogesterone caproate injection),Myalept (metreleptin for injection), Qnasl (beclomethasone dipropionate)nasal aerosol, Simponi (golimumab), Sitavig (acyclovir) buccal tablets,Tecfidera (dimethyl fumarate), Tivicay (dolutegravir), VariZIG,Varicella Zoster Immune Globulin (Human), Flublok (seasonal influenzavaccine), Flucelvax (influenza virus vaccine), Fulyzaq (crofelemer),Horizant (gabapentin enacarbil), Qnasl (beclomethasone dipropionate)nasal aerosol, Rayos (prednisone) delayed-release tablets, Stribild(elvitegravir, cobicistat, emtricitabine, tenofovir disoproxilfumarate), Tudorza Pressair (aclidinium bromide inhalation powder),Arcapta (indacaterol maleate inhalation powder), Benlysta (belimumab),Complera (emtricitabine/rilpivirine/tenofovir disoproxil fumarate),Daliresp (roflumilast), Dificid (fidaxomicin), Edurant (rilpivirine),Firazyr (icatibant), Gralise (gabapentin), Incivek (telaprevir), Nulojix(belatacept), Victrelis (boceprevir), Cayston (aztreonam for inhalationsolution), Egrifta (tesamorelin for injection), Menveo (meningitisvaccine), Oravig (miconazole), Prevnar 13 (Pneumococcal 13-valentConjugate Vaccine), Teflaro (ceftaroline fosamil), Zortress(everolimus), Zymaxid (gatifloxacin ophthalmic solution), Bepreve(bepotastine besilate ophthalmic solution), Berinert (C1 EsteraseInhibitor (Human)), Besivance (besifloxacin ophthalmic suspension),Cervarix [Human Papillomavirus Bivalent (Types 16 and 18) Vaccine,Recombinant], Coartem (artemether/lumefantrine), Hiberix (Haemophilus bConjugate Vaccine; Tetanus Toxoid Conjugate), Ilaris (canakinumab),Ixiaro (Japanese Encephalitis Vaccine, Inactivated, Adsorbed), Kalbitor(ecallantide), Qutenza (capsaicin), Vibativ (telavancin), Zirgan(ganciclovir ophthalmic gel), Aptivus (tipranavir), Astepro (azelastinehydrochloride nasal spray), Cinryze (C1 Inhibitor (Human)), Intelence(etravirine), Moxatag (amoxicillin), Rotarix (Rotavirus Vaccine, Live,Oral), Tysabri (natalizumab), Viread (tenofovir disoproxil fumarate),Altabax (retapamulin), AzaSite (azithromycin), Doribax (doripenem),Extina (ketoconazole), Isentress (raltegravir), Selzentry (maraviroc),Veramyst (fluticasone furoate), Xyzal (levocetirizine dihydrochloride),Eraxis (anidulafungin), Gardasil (quadrivalent human papillomavirus(types 6, 11, 16, 18) recombinant vaccine), Noxafil (posaconazole),Prezista (darunavir), Rotateq (rotavirus vaccine, live oralpentavalent), Tyzeka (telbivudine), Veregen (kunecatechins), Baraclude(entecavir), Tygacil (tigecycline), Ketek (telithromycin), Tindamax,tinidazole, Xifaxan (rifaximin), Amevive (alefacept), FluMist (InfluenzaVirus Vaccine), Fuzeon (enfuvirtide), Lexiva (fosamprenavir calcium),Reyataz (atazanavir sulfate), Alinia (nitazoxanide), Clarinex, Daptacel,Fluzone Preservative-free, Hepsera (adefovir dipivoxil), PediarixVaccine, Pegasys (peginterferon alfa-2a), Restasis (cyclosporineophthalmic emulsion), Sustiva, Vfend (voriconazole), Avelox I.V.(moxifloxacin hydrochloride), Cancidas, Peg-Intron (peginterferonalfa-2b), Rebetol (ribavirin), Spectracef, Twinrix, Valcyte(valganciclovir HCl), Xigris (drotrecogin alfa [activated]), ABREVA(docosanol), Biaxin XL (clarithromycin extended-release tablets),Cefazolin and Dextrose USP, Children's Motrin Cold, Evoxac, KaletraCapsules and Oral Solution, Lamisil (terbinafine hydrochloride) Solution(1%), Lotrisone (clotrimazole/betamethasone diproprionate) lotion,Malarone (atovaquone; proguanil hydrochloride) Tablet, Rapamune(sirolimus) Tablets, Rid Mousse, Tri-Nasal Spray (triamcinoloneacetonide spray), Trivagizole 3 (clotrimazole) Vaginal Cream, Trizivir(abacavir sulfate; lamivudine; zidovudine AZT) Tablet, Agenerase(amprenavir), Cleocin (clindamycin phosphate), Famvir (famciclovir),Norvir (ritonavir), Panretin Gel, Rapamune (sirolimus) oral solution,Relenza, Synercid I.V., Tamiflu capsule, Vistide (cidofovir), Allegra-D,CellCept, Clemastine fumarate syrup, Dynabac, REBETRON™ CombinationTherapy, Simulect, Timentin, Viroptic, INFANRIX (Diphtheria and TetanusToxoids and Acellular Pertussis Vaccine Adsorbed), Acyclovir Capsules,Aldara (imiquimod), Aphthasol, Combivir, Condylox Gel 0.5% (pokofilox),Flagyl ER, Flonase Nasal Spray, Fortovase, INFERGEN (interferonalfacon-1), Intron A (interferon alfa-2b, recombinant), RescriptorTablets (delavirdine mesylate tablets), SPORANOX (itraconazole),Stromectol (ivermectin), Taxol, Trovan, VIRACEPT (nelfinavir mesylate),Zerit (stavudine), Albenza (albendazole), Apthasol (Amlexanox),Carrington patch, Confide, Crixivan (Indinavir sulfate), Gastrocrom OralConcentrate (cromolyn sodium), Havrix, Lamisil (terbinafinehydrochloride) Tablets, Leukine (sargramostim), Oral Cytovene, RespiGam(Respiratory Syncitial Virus Immune Globulin Intravenous), Videx(didanosine), Viramune (nevirapine), Vitrasert Implant, Zithromax(azithromycin), Cedax (ceftibuten), Clarithromycin (Biaxin), Epivir(lamivudine), Invirase (saquinavir), Valtrex (valacyclovir HCl), andZyrtec (cetirizine HCl).

Further therapeutic agents that can be combined with a subject compoundcan be found in Goodman and Gilman's “The Pharmacological Basis ofTherapeutics” Tenth Edition edited by Hardman, Limbird and Gilman or thePhysician's Desk Reference, both of which are incorporated herein byreference in their entirety.

The compounds described herein can be used in combination with theagents provided herein or other suitable agents, depending on thecondition being treated. Hence, in some embodiments, the compounds asprovided herein will be co-administered with other agents as describedabove. When used in combination therapy, the compounds described hereincan be administered with the second agent simultaneously or separately.This administration in combination can include simultaneousadministration of the two agents in the same dosage form, simultaneousadministration in separate dosage forms, and separate administration.That is, a compound described herein and any of the agents describedabove can be formulated together in the same dosage form andadministered simultaneously. Alternatively, a compound as providedherein and any of the agents described above can be simultaneouslyadministered, wherein both the agents are present in separateformulations. In another alternative, a compound as provided herein canbe administered just followed by and any of the agents described above,or vice versa. In the separate administration protocol, a compound asprovided herein and any of the agents described above can beadministered a few minutes apart, or a few hours apart, or a few daysapart.

Administration of the compounds as provided herein can be effected byany method that enables delivery of the compounds to the site of action.An effective amount of a compound as provided herein can be administeredin either single or multiple doses by any of the accepted modes ofadministration of agents having similar utilities, including rectal,buccal, intranasal and transdermal routes, by intra-arterial injection,intravenously, intraperitoneally, parenterally, intramuscularly,subcutaneously, orally, topically, as an inhalant, or via an impregnatedor coated device such as a stent, for example, or an artery-insertedcylindrical polymer.

When a compound as provided herein is administered in a pharmaceuticalcomposition that comprises one or more agents, and the agent has ashorter half-life than the compound as provided herein, unit dose formsof the agent and the compound as provided herein can be adjustedaccordingly.

6. EXAMPLES

The examples and preparations provided below further illustrate andexemplify the compounds as provided herein and methods of preparing suchcompounds. It is to be understood that the scope of the presentdisclosure is not limited in any way by the scope of the followingexamples and preparations. In the following examples molecules with asingle chiral center, unless otherwise noted, exist as a racemicmixture. Those molecules with two or more chiral centers, unlessotherwise noted, exist as a racemic mixture of diastereomers. Singleenantiomers/diastereomers can be obtained by methods known to thoseskilled in the art.

Abbreviations/Acronyms Full Name/Description ACN or MeCN acetonitrileDCM dichloromethane DMF dimethylformamide EtOAc ethyl acetate IPAisopropyl alcohol IPAc isopropyl acetate MEK methyl ethyl ketone 2-MeTHF2-methyltetrahydroruran MIBK methyl iso-butyl ketone MTBE or TBMEtert-butyl methyl ether THF tetrahydrofuran DIPEA diisopropylethylamineDIPA Diisopropanolamine EDCI 3-(3-Dimethylaminopropyl)-1-ethyl-carbodiimide hydrochloride

Example 1 Preparation of Compound 1

A. Method 1

Compound 1 was prepared in 3 steps from compound A according to thefollowing procedures: Compound A was coupled to2-((tert-butoxycarbonyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid according to the following procedure: Compound A (27.4 mmol, 1.0equiv), HOBt hydrate (1.2 equiv),2-((tert-butoxycarbonyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (1.05 equiv) and EDCI (1.25 equiv) were added to a 200 mL roundbottomed flask with a stir bar. N,N-Dimethylformamide (50 mL) was addedand the suspension was stirred at RT for 2 min. Hunig's base (4.0 equiv)was added and after which the suspension became homogeneous and wasstirred for 22 h resulting in the formation of a solid cake in thereaction flask. The solid mixture was added to water (600 mL) andstirred for 3 h. The resulting cream colored solid was filtered andwashed with water (2×100 mL) and dried. The solid was then dissolved inmethylene chloride (40 mL) after which trifluoroacetic acid (10 equiv,20 mL) was added and the reaction was stirred for 30 min at RT afterwhich there is no more starting material by LC/MS analysis. The solutionwas then concentrated and coevaporated with a mixture of methylenechloride/ethanol (1:1 v/v) and then dried under high vacuum overnight.The resulting solid was triturated with 60 mL of ethanol for 1 h andthen collected via vacuum filtration. The beige solid was thenneutralized with sodium carbonate solution (100 mL) and then transferredto a separatory funnel with methylene chloride (350 mL). The water layerwas extracted with an additional 100 mL of methylene chloride. Thecombined organic layers were dried over sodium sulfate, filtered andconcentrated under vacuum to provide a pale yellow solid that waspurified using flash silica gel chromatography (Combiflash, 24 g column,gradient of 0-5% methanol/methylene chloride) to provide amide B. ESI-MSm/z: 459.4 [M+H]+.

Amide B was placed in a sealed tube (0.67 mmol, 1.0 equiv) followed bydichlorobis(acetonitrile)palladium (15 mol %), X-Phos (45 mol %), andcesium carbonate (3.0 equiv) Propionitrile (5 mL) was added and themixture was bubbled with Ar for 1 min. 4-Ethynyl-1-methyl-1H-pyrazole(1.24 equiv) was added and the resulting orange mixture was sealed andstirred in an oil bath at 85° C. for 1.5 h. The resulting brownish-blackmixture was allowed to cool at which point there was no more SM by LC/MSanalysis. The mixture was then filtered through a short plug of cottonusing acetonitrile and methylene chloride. The combined filtrates wereconcentrated onto silica gel and purified using flash silica gelchromatography (Combiflash, 4 g column, gradient of 0-5% methylenechloride/methanol). The resulting material was further purified byreverse phase HPLC (15-90% acetonitrile with 0.1% formic acid/water with0.1% formic water) to provide desired Compound 1. The solutioncontaining Compound 1 was lyophilized to give amorphous Compound 1.ESI-MS m/z: 529.5 [M+H]+.

B. Method 2

The first step started with palladium-catalyzed cross-coupling ofCompound A with 4-ethynyl-1-methyl-1H-pyrazole in acetonitrile to formCompound C. The allowable temperature range for this coupling was 70±5°C. Once the cross-coupling reaction was complete, the reaction mixturewas cooled to room temperature and filtered to remove insolubleinorganic salts. The resulting solution was first treated with afunctionalized silica gel followed by regular silica gel. Crude CompoundC was collected by filtration and washing of the silica gel with a 30%propyl acetate/acetonitrile solution. Crystallization of Compound C wasachieved by solvent exchange into n-PrOAc and crystals were isolated byfiltration and drying to within specification limits. Typical chemicalpurity is >98% area. Typical yields are 70-80%.

More specifically, acetonitrile (ACS grade, MeCN) was degassed withnitrogen for at least 30 minutes. Degassed acetonitrile (2 volumes)under nitrogen was charged to the reaction vessel. Compound A (1 wt, 1equ.), dichlorobis(acetonitrile)palladium (II) (0.05 equ.),dicyclohexyl[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (XPhos)(0.1025 equ.) and cesium carbonate (1.2 equ.) under nitrogen were alsocharged to the reaction vessel. The reaction mixture was agitated.4-ethynyl-1-methyl-1H-pyrazole (1.2 equ. corrected for assay content)was charged into the reaction vessel. Heat the reaction mixture to 70±2°C. (internal temperature).

The reaction was monitored by HPLC after 3 hours. Once the reaction wasdetermined complete the heat was turned off and the reaction mixturecooled to room temperature (22±3° C.) over a minimum of 2 hours and heldat r.t. for ≥2 hours. The suspension was filtered and the cake washedwith acetonitrile (2×2 volumes). The washes were combined with thefiltrate (product layer). The filtrate (product layer) was transferredback into the reaction vessel. The vessel was flushed with nitrogen forat least 2 minutes and agitation re-started. 2-mercaptoethyl ethylsulfide Silica [PhosphonicS SEM26] (0.5 wt % with respect to Compound A)was charged at room temperature (22±3° C.). The mixture was stirred for≥4 hours at r.t. The resin was filtered off and washed with acetonitrile(2×2 volumes). The washes were combined with the filtrate (productlayer).

The acetonitrile solution (product layer) was pumped through apre-packed silica cartridge (flow rate≈50 mL/min) and collected thefiltrate in a clean vessel. The silica column was washed with 30% propylacetate in acetonitrile (5×4 volumes). The washes were combined with theprevious filtrate. The acetonitrile/propyl acetate solution wastransferred into a clean distillation vessel. The volume was reduced to10 volumes under vacuum (200 Torr, 45° C.). Propyl acetate (10 volumes)was charged to the distillation vessel. The reaction volume was reducedto 10 volumes under vacuum (140 Torr, 50° C.). The vacuum was releasedand propyl acetate (10 volumes) was charged to the distillation vessel.The reaction volume was reduced to 10 volumes under vacuum (100 Torr,50° C.).

The PrOAc solution/mixture was heated to reflux (Target 102° C.) withstirring, under nitrogen, and was held for ≥30 minutes. The heat wasturned off and the mixture was cooled to r.t. (22±3° C.) over 4 hours.Heptane (10 volumes, ACS grade) was charged to the suspension at r.t.(22±3° C.) over 30 minutes. The mixture was aged for ≥1 hour at r.t.(22±3° C.). The solid was filtered off and washed with heptane (2×5volumes). The solid was deliquored under vacuum for ≥2 hours. The solidwas transferred to drying trays and placed in a vacuum oven and dried at40° C. under vacuum, with a nitrogen bleed, until constant weight wasachieved.

The second step is an amide coupling reaction between Compound C and thecarboxylic acid employing EDCI and HOBt at 30° C. Compound C was treatedwith the carboxylic acid, HOBt, EDCI and DIPEA in DMF and stirred at 30°C. under nitrogen until judged complete by HPLC. The reaction mixturewas polish filtered and pumped slowly into a 0.1M potassium carbonateaq. solution. After ageing, the resulting suspension was filtered. Thecrude Compound 1 was subsequently purified by recrystallization. Thesolid was dissolved in a mixture of hot IPA/water (1:1) and treated withwater. On cooling Compound 1 precipitated from solution. The crystalswere isolated by filtration, washed with water and dried. Typicalchemical purity is >98% area. Typical yields are 60-70%.

Specifically, the reaction vessel was charged with Compound C (1 wt, 1equ.), 2-aminopyrazolo[1,5-a]pyrimidinecarboxylic acid (1.05 equ),1-hydroxybenzotriazole hydrate (0.1 equ.), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, anhydrous (1.2 equ.),N,N-dimethylformamide, ACS grade (10 volumes), andN,N-diisopropylethylamine, ReagentPlus (3 equ.) at r.t. (22±3° C.) withunder nitrogen at r.t. The reaction mixture was heated at 30±5° C. undernitrogen. The reaction was stirred at 30±5° C. (Target 30° C.). Thereaction was monitored by HPLC after 16 hours. This reaction could alsobe conducted without the base, N-diisopropylethylamine.

The reaction mixture was quenched as follows. A 0.1M solution ofpotassium carbonate in water was prepared. The quench vessel was chargedwith potassium carbonate (3.2 equ) followed by water (80 volumes). Themixture was stirred at r.t. until fully dissolved. The DMF reactionsolution (product layer) was polish filtered. The reaction vessel wasrinsed with a small amount of DMF (0.5 volumes). The DMF rinse waspassed through the polish filter. The DMF solution (product layer) wasslowly transferred into the potassium carbonate solution over ≥2 hoursat r.t (22±3° C.). The suspension was aged for ≥1 hour at r.t. (22±3°C.). The solid was filtered off under house vacuum (table top filter),de-liquored and washed with water (2×10 volumes). It was deliquoredunder vacuum at r.t. until constant weight was achieved.

A vessel was charged with crude Compound 1, isopropyl alcohol (10volumes) followed by water (10 volumes). The mixture was agitated. Thesuspension was heated to 80±3° C. (Target 80° C.). The mixture was agedfor ≥15 minutes. Water (10 volumes) was charged to the vessel at such arate to maintain the internal temperature between 80±5° C. (Target 80°C.). The internal temperature was adjusted to 80° C. and held for ≥20minutes before proceeding to the next step. The suspension was cooled tor.t. (22±3° C.) over 4 hours. The suspension was aged at r.t. for ≥4hours. The solid was filtered under vacuum. The solid was washed with20% isopropyl alcohol in water (2×5 volumes). The solid was deliquoredunder vacuum for ≥1 hour. The solid was transferred to drying trays andplaced in a vacuum oven and dry at 40° C. under vacuum, with a nitrogenbleed, until constant weight was achieved. Characterization of the solidshowed it was Form 1.

C. Method 3

The first step is palladium-catalyzed cross-coupling of Compound A with4-ethynyl-1-methyl-1H-pyrazole in acetonitrile to form Compound C. Thesame procedures from Method 2 above can be used to produce compound C.Compound C was coupled with compound D to form Compound 1.

Compound D was prepared by converting the carboxylic acid to its NHSactivated ester.

The reaction vessel was flushed with nitrogen for ≥15 minutes. DMF (2volumes) was charged to the vessel. The jacket temperature was set to23° C. N-Hydroxysuccinimide (HOSu or NHS) (1.3 equ.) was charged to thereaction vessel, followed by 2-aminopyrazolo[1,5-a]pyrimidinecarboxylicacid (1 wt, 1 equ.) and EDCI (1.3 equ.) and DMF (13 volumes) at 23° C.under nitrogen.

The reaction mixture was stirred at 23° C. under nitrogen. The reactionwas monitored by HPLC. Once judged complete the reaction contents werecooled to 0±5° C. Water (20 volumes) was added over ≥4 hours to thereaction mixture via peristaltic pump) under nitrogen at such a rate tomaintain the internal temperature at 0±5° C. The suspension was aged for1 hour at 0±5° C. The solid was filtered off under vacuum and washedwith water (2×5 volumes). The solid was deliquored for ≥1 hour thentransferred into a vacuum oven and dried at 40° C., under vacuum, with aslight nitrogen purge until constant weight was achieved. In analternative example, the reaction was ran without DIPEA.

The coupling of compound C with compound D was carried out as follows.The reaction vessel was evacuated and purged with nitrogen for ≥15minutes. Compound C (1 wt, 1 equ.), Compound D (1.05 equ.) andacetonitrile (15 volumes), were charged to the reaction vessel at 22±3°C., under nitrogen. The reaction vessel was evacuated and back filledwith nitrogen. The mixture was agitated. N,N-Diisopropylethylamine(DIPEA) (1.1 equ.) at 22±3° C. was charged to the reaction vessel undernitrogen. The reaction mixture was heated to 60±2° C. and stirred undernitrogen. The reaction was monitored by HPLC.

Once the reaction was judged complete by HPLC a 1M solution of potassiumhydroxide (0.5 equ., 1.21 volumes) was charged to the reaction vessel atsuch a rate to maintain an internal temperature between 52-60° C. Thesuspension was aged at 60±2° C. for 2 hours, then cooled to 22±3° C.over 1 hour. The solid was filtered off under vacuum then washed withacetonitrile (2×2.5 volumes) followed by 4:1/water: acetonitrile (2×2.5volumes). It was deliquored under vacuum at r.t. for ≥12 hours.

Water (8 volumes), crude compound 1 (1 wt) from and acetone (32 volumes)were charged to the recrystallization vessel. The mixture was agitatedand heated to 55±5° C. The solution was aged at 55±5° C. (Target 50° C.)for 10 minutes and then cooled the solution to 23±3° C. The reactionsolution was transferred into a holding drum. The recrystallizationvessel was rinsed with acetone (twice). The reflux condenser wasexchanged for a short path distillation setup on the recrystallizationvessel and flushed with nitrogen for a minimum of 15 minutes. Thereaction solution was transferred through an inline polish filter (mediaPTFE, pore size 1 μm) back into the recrystallization vessel. Acetone (2volumes) was charged into the holding drum and the rinses weretransferred into the recrystallization vessel through the inline filter.The mixture was agitated and the jacket temperature was set to 45° C.

The solvent volume was reduced to 20 volumes under reduced pressuremaintaining an internal temperature at ≥30° C. (Target 35° C.). Thevacuum was released and a mixture of 3:2/isopropyl alcohol:water (20volumes) was charged to the recrystallization vessel. The solvent volumewas reduced to 20 volumes under reduced pressure maintaining an internaltemperature at ≥30° C. (Target 35° C.). The vacuum was released and amixture of 3:2/isopropyl alcohol:water (20 volumes) was charged to therecrystallization vessel. The solvent volume was reduced to 20 volumesunder reduced pressure maintaining an internal temperature at ≥30° C.(Target 35° C.). The vacuum was released and a mixture of 3:2/isopropylalcohol:water (10 volumes) was charged to the recrystallization vessel.The short path distillation head was replaced with a reflux condenser.

The reactor was purged with nitrogen for at least 15 minutes. Thestirred suspension was heated to 60±2° C. (Target 60° C.). Thesuspension was aged at 60±2° C. (Target 60° C.) for 14±2 hours and wasthen cooled to 23±3° C. over ≥1 hour. A small sample was taken toconfirm conversion to Form 1 was complete. The suspension was aged for≥3 hours. The solid was filtered off under vacuum and washed the solidwith 20% isopropyl alcohol in water (2×5 volumes). The solid wasdeliquored under vacuum for ≥1 hour, transferred to drying trays, placedin a vacuum oven and dried at 40° C. under vacuum, with a nitrogenbleed, until constant weight was achieved. Characterization of the solidshowed it was Form 1.

The typical yield for Method 3 is about 75-85%.

Example 2 Polymorph Screen

A. Solubility Screen

Amorphous Compound 1 (30 mg) was weighed out and 100 μl (˜3 vols)solvent was added at room temperature. Many samples initially dissolvedbut precipitated back out again after stirring for a few minutes. It wasassumed that these samples had crystallized, so additional portions ofsolvent were added to determine the solubility of the crystallinematerial. See Table 1 for solubility assessment of amorphous Compound 1.

Samples were stirred at 50° C. for 1 hour then cooled to 5° C. using alinear cooling rate of 0.1° C./min. Samples were stirred at 5° C. for ˜9hours then observations were made and small portions of the samplescontaining solids were filtered, air dried and analyzed by XRPD.

Crystalline samples were filtered and dried in a vacuum oven for 3 daysat RT prior to characterization. Amorphous samples were matured (shakenin cycles of 4 hours at RT/4 hours at 50° C.) and were analyzed by XRPDafter 32 days. Solutions were stored in the freezer, and then left toevaporate at ambient if no precipitate formed at −20° C. Solids producedby evaporation were analyzed by XRPD.

B. Maturation Screen

This screen was carried out using all the solvents from the solubilityscreen except DMF (as it was too soluble to make a slurry) and thosesolvents that were very poorly soluble (as suspension in these solventsfrom the solubility screen were already maturing). Anisole was added asan additional solvent for this screen.

Amorphous Compound 1 (30 mg) was prepared in glass vials and 500 μl (˜17vols) solvent was added at 25° C. The samples were shaken in thematuration chamber (cycles of 4 hours at RT/4 hours at 50° C.) for 3days. Small aliquots of samples containing solids were filtered andanalyzed by XRPD. Samples containing crystals were analyzed by opticalmicroscopy to assess suitability for SXRD determination prior to XRPDanalysis. Solutions were left to evaporate at ambient conditions thenany resulting solids were analyzed by XRPD. Selected solids werefiltered and dried in a vacuum oven for 2 days prior tocharacterization.

C. Low Temperature Screen

This screen was carried out with the same set of solvents as were usedin the solubility screen and one additional solvent (anisole).

Amorphous Compound 1 (30 mg) was prepared in glass vials and cooled to5° C. Solvent (either 500 μl (˜17 vols) or 250 μl (˜8 vols) based on theresults from solubility screen) was added and samples were stirred at 5°C. overnight. Small aliquots of samples containing solids were filteredand analyzed by XRPD. Solutions were left to evaporate at ambientconditions then any resulting solids were analyzed by XRPD. Selectedsolids were filtered and dried in a vacuum oven for 2 days prior tocharacterization. Samples containing poorly crystalline solids were leftto stir at 5° C. for a total of 40 days and then were re-analyzed byXRPD.

D. Maturation Screen—Mixed Solvents

Amorphous Compound 1 (30 mg) was prepared in glass vials and 500 μl (˜17vols) of mixed solvents (pre-mixed using 50/50 v/v) was added at roomtemperature. Samples were matured (shaken in cycles of 4 hours at 25°C./4 hours at 50° C.) for 2 days. Small aliquots of samples containingsolids were filtered and analyzed by XRPD. Solutions were left toevaporate at ambient conditions then any resulting solids were analyzedby XRPD.

E. Results

Table 1 summarizes the solubility assessment of amorphous Compound 1.During the initial solubility assessment of the amorphous material itwas observed that many samples initially dissolved but precipitated backout again after stirring for a few minutes. It was assumed that thesehad precipitated as a crystalline form of the free base. Further solventwas added to assess the solubility of the crystalline form, which wasfound to be much less soluble than the amorphous material.

The results of the polymorph screen are summarized in Table 2. Theresults of the maturation screen in mixed solvents are summarized inTable 3. The polymorph screens showed a total of six distinct forms ofthe free base. Forms 1, 2, 3 and 4 were crystallized from a range ofsolvents. Forms 5 and 6 were made by maturing in anisole and byslurrying at low temperatures in nitromethane, respectively. All samplesthat displayed good crystallinity were filtered and dried before beinganalyzed by high resolution XRPD.

Table 4 provides the results of the damp and dried samples of varioussamples. Table 5 provides further characterization results of thevarious forms of Compound 1.

Forms 2, 3 and 4 were found to be a set of pseudo-isostructuralsolvates. The XRPD diffractograms are grouped into Forms 2, 3 and 4 butwithin each group there are small differences in the diffractograms,indicating some variation in the crystal lattice due to the inclusion ofdifferent sized solvent molecules. A representative ORTEP (Oak RidgeThermal Ellipsoid Plot) plot of crystal structure of Form 2 is shown inFIG. 21, which shows that there is one molecule of solvent for eachmolecule of Compound 1. The same relationship may apply to all samplesof Forms 2, 3 and 4, e.g., that they are all mono-solvates, whichpartially de-solvate on drying. This is borne out by the residualsolvent levels seen in the ¹H NMR data (not shown), which are mostlyslightly less than 1 mole of solvent.

Most of the samples analyzed after drying were the same form as the dampsample, although some had reduced crystallinity. Where most of thesolvent was removed by drying, the samples showed a conversion to Form 7(a partially crystalline, partially solvated form).

The thermal behavior of all of these samples shows events relating tothe loss of solvent in the TGA and DSC followed by a melt, then are-crystallisation event. The re-crystallized sample was consistent withForm 1, showing that these solvates all convert to Form 1, once thesolvent has been removed by heating.

Form 5 was a meta-stable anisole solvate that converted to Form 1 ondrying in a vacuum oven. The characterization data for this dried samplewas consistent with Form 1.

Form 6 was made in the polymorph screen (from nitromethane) and was alsomade by sonication in the cocrystal experiments provided herein (fromACN/water 50/50 v/v). The sample showed no presence of acetonitrile orconformer. This form is a hydrated form of Compound 1. Using the weightloss seen in the TGA experiment this form is a tri-hydrate form of thefree base.

Representative XRPD, TGA, DSC, and GVS characterizations of Forms 1-7are shown in FIGS. 1-19.

TABLE 1 Solubility assessment of amorphous Compound 1 Solvent Solubilityn-Heptane <17 mg/ml Ethyl acetate <17 mg/ml Isopropyl acetate <17 mg/mlMIBK <17 mg/ml 2-Propanol <17 mg/ml MEK <17 mg/ml 1-Propanol <17 mg/mlAcetone <17 mg/ml Ethanol <17 mg/ml Dimethyl sulfoxide >=333 mg/ml Water<17 mg/ml tert-Butylmethyl ether <17 mg/ml 2-Methyl-1-propanol <17 mg/mlCyclohexane <17 mg/ml 1,4-Dioxane <17 mg/ml Toluene <17 mg/ml Chloroform<17 mg/ml 1,2-Dimethoxyethane <17 mg/ml Tetrahydrofuran <17 mg/mlDichloromethane 17-20 mg/ml 2-Methoxyethanol >=333 mg/ml Methanol <17mg/ml N,N-Dimethylformamide >=333 mg/ml Acetonitrile <17 mg/mlEthyleneglycol <17 mg/ml Nitromethane 25-33 mg/ml THF/Water (95/5 v/v)<17 mg/ml IPA/Water (95/5 v/v) <17 mg/ml

TABLE 2 Summary of results from the polymorph screens Solubility screenMaturation screen Low temperature screen after maturation (mat) afterevaporation after evaporation Solvent initial result or evaporation(evap) (evap) (evap) n-Heptane amorphous crystalline, Form 1 n/aamorphous (mat) Ethyl acetate partially crystalline, partiallycrystalline, n/a crystalline Form 1 Form 1 Form 1 (mat) Isopropylacetate partially crystalline, amorphous (mat) n/a crystalline Form 1Form 1 MIBK crystalline, Form 1 n/a crystalline, Form 1 crystalline Form1 2-Propanol crystalline crystalline Form 4 crystalline, Form 4crystalline Form 4 (mat) MEK crystalline Form 3 n/a crystalline, Form 3crystalline Form 3 1-Propanol crystalline Form 3 + n/a crystalline, Form2 crystalline Form 4 extra peaks Acetone crystalline, similar n/acrystalline, Form 2 crystalline Form 4 to Form 3 Ethanol crystalline,Form 1 n/a partially crystalline, partially crystalline Form 1 Form 1Dimethyl crystalline Form 4 n/a crystalline, Form 4 crystalline Form 3sulfoxide (evap) Water amorphous crystalline, Form 1 n/a amorphous (mat)tert-Butylmethyl crystalline, Form 3 n/a crystalline, Form 3 crystallineForm 3 ether 2-Methyl-1- crystalline, Form 3 crystalline Form 4crystalline, Form 3 crystalline Form 4 propanol (mat) Cyclohexaneamorphous crystalline, Form 1 n/a amorphous (mat) 1,4-Dioxane partiallycrystalline, solution (mat), crystalline, Form 4 crystalline Form 4 Form4 oil (evap) Toluene crystalline, Form 1 n/a crystalline, Form 1crystalline Form 1 Chloroform crystalline, Form 2 n/a crystalline, Form4 partially crystalline Form 4 1,2- crystalline, Form 1 n/a crystalline,Form 1 crystalline Form 1 Dimethoxyethane Tetrahydrofuran crystalline,Form 4 n/a crystalline, Form 4 crystalline Form 4 Dichloromethanesolution crystalline, Form 2 crystalline, Form 2 crystalline Form 2(evap) (evap) 2-Methoxyethanol crystalline, Form 1 n/a weaklycrystalline, crystalline Form 4 Form 1 (evap) Methanol crystalline, Form1 n/a crystalline, Form 1 partially crystalline Form 1 N,N- crystalline,Form 1 + n/a n/a crystalline, Form 1 Dimethylformamide extra peak (evap)Acetonitrile crystalline, Form 1 n/a crystalline, Form 1 crystallineForm 1 Ethyleneglycol crystalline crystalline, Form 1 weaklycrystalline, crystalline Form 1 (mat) Form 1 Nitromethane solutioncrystalline, Form 1 weakly crystalline, crystalline Form 6 (evap) Form 1(evap) THF/Water crystalline, Form 4 n/a crystalline, Form 4 crystallineForm 4 (95/5 v/v) IPA/Water partially crystalline, crystalline Form 4crystalline, Form 4 crystalline Form 4 (95/5 v/v) Form 4 (mat) Anisolen/a n/a crystalline, Form 5 partially crystalline Form 1

TABLE 3 Summary of results from polymorph screen with mixed solventsSolvent mixture (mixtures are 50/50 v/v) XRPD results 2-Methyl THFcrystalline Form 3 2-Methyl THF/IPA crystalline Form 3 2-MethylTHF/Ethanol crystalline Form 3 + extra peak 2-Methyl THF/Toluenecrystalline Form 3 Anisole/IPA crystalline Form 1 Anisole/Ethanolcrystalline Form 1 Anisole/Toluene crystalline Form 1 Anisole/THFcrystalline Form 4 Acetonitrile/Water crystalline Form 1 Toluene/THFcrystalline Form 4 Toluene/Ethanol crystalline Form 1 Toluene/IPAcrystalline Form 4 DCM/Acetone crystalline Form 2

TABLE 4 Damp and dry screening samples of Compound 1 XRPD XRPD of dampof dried Method Solvent sample sample maturation 1-propanol Form 2 Form2 solubility screen chloroform Form 2 Form 2 solubility screen DCM Form2 Form 2 maturation 2-methyl-1-propanol Form 3 Form 3 high temperatureMEK Form 3 partially crystalline new phase Form 7 solubility screen1-propanol Form 3 weakly crystalline Form 7 solubility screen acetoneForm 3 weakly crystalline Form 7 solubility screen TBME Form 3 partiallycrystalline Form 7 maturation 2-propanol Form 4 Form 4 maturationIPA/water 95/5 v/v Form 4 Form 4 solubility screen DMSO Form 4 Form 4solubility screen THF Form 4 weakly crystalline Form 4 solubility screenTHF/water 95/5 v/v Form 4 weakly crystalline Form 4 maturationchloroform Form 4 partially crystalline Form 4 maturation THF/water 95/5v/v Form 4 some changes, similar to Form 2 low temperature 1-propanolForm 4 partially crystalline Form 4 low temperature acetone Form 4weakly crystalline Form 7 low temperature 2-methyl-1-propanol Form 4weakly crystalline Form 4 low temperature 1,4-dioxane Form 4 weaklycrystalline Form 4 low temperature 2-methoxyethanol Form 4 almostamorphous (insufficient sample) maturation anisole Form 5 changes toForm 1 low temperature nitromethane Form 6 inconclusive - insufficientsample sonication in ACN/water Form 6 crystalline, Form 6 cocrystalscreen 50/50 v/v

TABLE 5 Characterization of crystalline forms of Compound 1 XRPD of XRPDof Purity damp dried by Static Stability Method Solvent sample sampleNMR HPLC TGA DSC 40° C./75% RH maturation 1-propanol Form 2 Form 2 0.85mole eq 99.3 0.9 wt % loss broad endotherm onset 25° C. (5.4 J/g)amorphous after 7 1-propanol area % 25-75° C. sharp endotherm onset 151°C. (57.9 J/g) and 36 days (8.7 wt %) 8.5 wt % loss broad exotherm onset179° C. (29.7 J/g) 75-250° C. sharp endotherm onset 244° C. (75.5 J/g)maturation 2-methyl- Form 3 Form 3 0.79 mole eq 99.2 0.8 wt % loss broadendotherm onset 29° C. (11.8 J/g) remains Form 3 1-propanol 2-methyl-1-area % 25-75° C. small endotherm onset 126° C. (0.9 J/g) but loss ofpropanol 10.4 wt % loss sharp endotherm onset 148° C. (54.5 J/g)crystallinity after (10.0 wt %) 75-300° C. broad exotherm onset 181° C.(26.9 J/g) 7 days and further sharp endotherm onset 246° C. (42.8 J/g)loss after 36 days high MEK Form 3 partially 0.25 mole eq 99.5 2.0 wt %loss broad endotherm onset 30° C. (21.6 J/g) remains Form 7 temper-crystalline MEK area % 25-80° C. sharp endotherm onset 127° C. (2.7 J/g)but loss of ature Form 7 (3.3 wt %) 3.4 wt % loss sharp endotherm onset137° C. (7.2 J/g) crystallinity after 80-175° C. broad exotherm onset169° C. (3.2 J/g) 7 days and further sharp endotherm onset 207° C. (1.3J/g) loss after 36 days broad endotherm onset 250° C. (7.7 J/g)maturation 2-propanol Form 4 Form 4 0.9 mole eq 99.0 0.7 wt % loss broadendotherm onset 30° C. (5.2 J/g) remains Form 4 IPA area % 25-75° C.sharp endotherm onset 156° C. (69.0 J/g) but loss of (9.3 wt %) 9.3 wt %loss broad exotherm onset 190° C. (28.0 J/g) crystallinity after 75-225°C. sharp endotherm onset 245° C. (43.7 J/g) 7 days and further lossafter 36 days maturation IPA/water Form 4 Form 4 0.83 mole eq 99.2 0.8wt % loss broad endotherm onset 28° C. (13.3 J/g) remains Form 4 95/5v/v IPA area % 25-75° C. sharp endotherm onset 156° C. (68.3 J/g) butloss of (8.6 wt %) 8.6 wt % loss broad exotherm onset 201° C. (17.2 J/g)crystallinity after 75-250° C. sharp endotherm onset 247° C. (29.1 J/g)7 days and further loss after 36 days maturation anisole Form 5 changesto 0.12 mole eq 99.0 no low small endotherm onset 126° C. (2.7 J/g) nochange after 7 Form 1 anisole area % temperature sharp endotherm onset254° C. (79.1 J/g) and 36 days (2.4 wt %) weight loss 2.1 wt % loss175-300° C. sonication ACN/ Form 6 Form 6 large water n/a 10.3% wt broadendotherm (onset 46° C., 274.0 J/g) n/a water peak but no loss 30-100°C. small endotherm (onset 154° C., 21.6 J/g) 50/50 v/v ACN or (3.3 moleeq small endotherm (onset 243° C., 4.9 J/g) coformer water)

Example 3 Preparation of Compound 1 Form 1 and Form 6

A. Preparation of Compound 1 Form 1

Exemplary methods to prepare Form 1 are described in Method 2 and Method3 of Example 1.

Another method to prepare Form 1 is described below. Form 1 wasscaled-up using ethanol Ethanol was chosen as a suitable ICH Class 3solvent. Amorphous Compound 1 (1 g) was prepared in a large glass vialand warmed to 50 C. Ethanol (17 ml, 17 vols) was added at 50° C. and thesample was stirred at 500 rpm. Initially the sample formed a gum butafter stirring for a few minutes, the gum converted to a bright yellowsuspension. The sample was stirred at 50° C. for 1 hour then an aliquotof the solid was filtered, air dried and analyzed by XRPD. The samplewas matured (shaken in cycles of 4 hours at 25° C./4 hours at 50° C.)for 2 days then another aliquot was filtered and analyzed by XRPD.

The remaining sample was allowed to cool to room temperature and wasfiltered through a 0.45 μm PTFE filter. The sample was air dried undervacuum for 1 hour and was dried in a vacuum oven overnight at 30° C. Theresulting solid, 930 mg (93 wt %), was analyzed by XRPD, ¹H NMR and byHPLC for purity. This method successfully produced crystalline Form 1material in a 93 wt % yield.

Another method to prepare Form 1 is described below. IPA-water (2 ml-2ml IPA-water) was mixed with amorphous Compound 1 (106 mg). Heat wasapplied to the sample until all Compound 1 is dissolved. The sample wasleft at room temperature overnight. Solid was collected and dried invacuum over at 45° C. for 2 hours. The XRPD shows that the material isForm 1.

B. Preparation of Compound 1 Form 6

Small-scale preparations of Form 6 were carried. Amorphous Compound 1(30 mg) was prepared in glass vials and cooled to 5° C. Solvent 1 wasadded (solvent and volume depend on the experiment, see Table 6) andobservations were made. Sample was stirred at 5° C. for a few secondsthen Solvent 2 was added (solvent and volume depend on the experiment,see Table 6) and further observations were made. Samples were stirred at5° C. for ˜10 minutes then aliquots of the sample were filtered, airdried and analyzed by XRPD. Samples were left to stir at 5° C. for afurther 2 hours then another set of aliquots were filtered, air driedand analyzed by XRPD. All experiments were carried out at 5° C.

TABLE 6 Summary of small scale experiments to identify a method to makeForm 6 Obs after Obs. On 10 mins Obs after 2 Obs. in addition ofstirring at hours Solvent 1 Vols solvent 1 solvent 2 Vols solvent 2 5°C. XRPD stirring at 5° C. XRPD 2- 3 vol solution water 20 suspension/suspension/ amorphous suspension amorphous methoxy vol gum gum (lessgum) ethanol nitromethane 3 vol solution water 20 emulsion/ gummycrystalline gummy crystalline vol gum solid Form 6 solid (more Form 6solid less gum) ACN 3 vol gum/ water 20 suspension/ suspension/amorphous suspension/ amorphous solution vol gum gum gum ACN 10 gum/water 20 suspension/ gummy partially suspension weakly vol solution volgum solid crystalline crystalline Form 6 Form 6 water 17 dry solid ACN17 suspension suspension crystalline suspension partially vol (floating)vol Form 6 crystalline Form 6 Obs.—observation

Form 6 material was successfully prepared in three of the fiveexperiments; the remaining experiments produced amorphous material. Thebest crystallinity was obtained from the experiments in nitromethane andwater/ACN (50/50 v/v). Results show that all the samples except thewater/ACN experiment formed gums (some of which converted to solids withstirring). Adding the water before the acetonitrile seems to prevent theformation of the gum, although the sample in water required vigorousshaking to wet the particles effectively. In addition, the crystallinityseemed to be worse after extended stirring. The best method for scale-upis the water/ACN (50/50 v/v) method with a short stir time beforeisolation.

A large scale-up to prepare Form 6 was carried out. Amorphous Compound 1(1 g) was prepared in a large glass vial and cooled to 5° C. Water (11.8ml, 12 vols) was added at 5° C. and the sample was shaken vigorously towet the particles. The sample was stirred at 500 rpm for a few minutesto form a suspension. Acetonitrile (7.2 ml, 7 vols) was added, initiallyforming a yellow gum. The sample was stirred for ˜5 minutes at 5° C.after which time the gum had converted to a powdery solid. The samplecan be seeded with a few milligrams of Form 6. An aliquot of the samplewas filtered, air dried and analyzed by XRPD. The sample was filteredthrough a 0.45 μm Nylon filter.

During filtration, a large lump of hard solid was found in thesuspension (from the gum). This solid was broken up with a spatula toproduce a powdery suspension and left to stir at 5° C. for a further 10minutes. An aliquot was taken and analyzed by XRPD. The remaining samplewas filtered through the same filter (added to the first crop) and thewhole sample was air dried under vacuum for 2.5 hours and was dried in avacuum oven at RT for 1 hour. The resulting solid, 796 mg (80 wt %), wasanalyzed by XRPD. This method successfully produced crystalline Form 6material in an 80 wt % yield. This method was repeated and Form 6 wasobtained in 87 wt % yield.

A comparison of the results for Forms 1 and 6 is shown in Table 7. TheForm 6 sample was found to be a hydrated form of the free base. Thewater content of the sample varied depending on the analysis and time.Two slightly different XRPD diffractograms were recorded approximately24 hours apart, between which the sample had been stored at ambientconditions.

The weight loss in the TGA and the water content by Karl Fischer do notagree exactly. These results can be explained by the GVS data. Theisotherm plot shows a large weight change, i.e. a large uptake/releaseof water between 20 and 60% RH. This means that at ambient conditionsthe water content of the sample will be very sensitive to the ambienthumidity. It seems that the water content is approximately 1-1.5 moleswater at ambient conditions (approximately 40% RH) but this is not afixed amount. The GVS results show a total change of approximately fivemoles of water between 0 and 90% RH; however, there are no plateaus inthe isotherm plot, which would indicate a zone of stability for anyparticular water content. Therefore, the water content is not stablewith respect to humidity. The varying water content also causes subtlechanges in the XRPD diffractogram.

TABLE 7 Characterization of Forms 1 and 6 of Compound 1 Sample Form 1reference Form 1 Form 6 Appearance white powder pale yellow powder paleyellow powder XRPD crystalline, Form 1 crystalline, Form 1 crystalline,Form 6 ¹H NMR consistent with structure, consistent with consistent withstructure, contains 0.05 mole IPA structure, contains contains noresidual solvent 0.29 mole ethanol but some water Purity by HPLC 98.3area % 99.1 area % 98.8 area % Optical tiny irregular particles in n/amixture of fused agglomerates microscopy agglomerates and primaryparticles. (approximately 1-5 μm) Particles are laths or irregularplates and are from <1 μm up to ~40 μm, average ~20 μm Karl Fischer n/an/a 4.9 wt % water (1.5 moles) TGA 0.6 wt % loss 230-310° C. n/a 4.1 wt% loss 25-90° C. (1.25 degradation after ~375° C. moles water)degradation after ~375° C. DSC sharp endotherm onset n/a broad endotherm(2 peaks) 255° C. (−85.7 J/g) onset 38° C. (−163.9 J/g), signs ofdegradation after consistent with weight loss 325° C. broad endothermonset 156° C. (−24.4 J/g) signs of degradation after 350° C. GVS totalweight change 0-90% n/a total weight change 0-90% RH RH ~0.5 wt %, non-15.0 wt % (5.2 moles water), hygroscopic. Steady hygroscopic. Largesteps 40-60% adsorption/desorption RH on adsorption and with nohysteresis. XRPD 40-20% RH on desorption analysis of the residue withsignificant hysteresis. showed no change in XRPD analysis of the residueform showed small changes Static Stability no change after 7 days, n/asubtle changes in XRPD after at 40° C./75% RH after 63 days no change 7days, still Form 6 in form but loss of no further changes after 26crystallinity days Static Stability no change after 7 days, n/a nochange in XRPD after 26 at 25° C./96% RH after 63 days no change days inform but loss of crystallinity

Example 4 Preparation of Compound 1 Form 8

A method of preparing Form 8 is described below. On heating, Form 6loses the water at relatively low temperatures and changes form to Form8. Form 8 is not consistent with the non-solvated Form 1 or with any ofthe other forms seen in the polymorph screens. It is likely to beanother non-solvated form of Compound 1. Further heating of Form 8 showsa melt endotherm in the DSC (onset 156° C., confirmed as a melt by VTX,data not shown). On cooling, the material remains amorphous. Arepresentative XRPD of Form Bis shown in FIG. 20.

Example 5 Competitive Slurry/Water Activity Experiments with Forms 1 and6

IPA/water and ACN/water were selected as suitable solvents for thecompetitive slurry/water activity experiments. These solvent mixtureswere selected to have some solubility of the Compound 1 free base and tobe able to obtain a wide range of water activities by varying the watercontent. IPA/water was selected as this solvent often formed solvatedforms of the free base and ACN/water was selected as this solventproduced only Form 1 of the free base during the polymorph screeningexperiments.

Results for the preparation of the saturated solutions are given inTable 8 and show that the amorphous material slurried in IPA/watermixtures crystallized to give the solvated Form 4. The ACN/watermixtures also showed that the amorphous material crystallized as Form 1in all conditions. There was one exception to this—the IPA/water sampleat 50° C. and at a water activity of 0.8 gave Form 1 rather than Form 4.The results of the competitive slurry experiments are given in Table 9.

TABLE 8 Experimental details and XRPD results from the preparation ofsaturated solutions Volume IPA/ Volume Water Temperature Solvent ACN(μl) water (μl) activity (° C.) Observation XRPD result IPA/water 980 200.2 5 suspension crystalline Form 4 IPA/water 980 20 0.2 25 suspensioncrystalline Form 4 IPA/water 980 20 0.2 50 suspension crystalline Form 4IPA/water 960 40 0.4 5 suspension crystalline Form 4 IPA/water 960 400.4 25 suspension crystalline Form 4 IPA/water 960 40 0.4 50 suspensioncrystalline Form 4 IPA/water 920 80 0.6 5 suspension crystalline Form 4IPA/water 920 80 0.6 25 suspension crystalline Form 4 IPA/water 920 800.6 50 suspension crystalline Form 4 IPA/water 840 160 0.8 5 suspensioncrystalline Form 4 IPA/water 840 160 0.8 25 suspension crystalline Form4 IPA/water 840 160 0.8 50 suspension crystalline Form 1 ACN/water 99010 0.2 5 suspension crystalline Form 1 ACN/water 990 10 0.2 25suspension crystalline Form 1 ACN/water 990 10 0.2 50 suspensioncrystalline Form 1 ACN/water 980 20 0.4 5 suspension crystalline Form 1ACN/water 980 20 0.4 25 suspension crystalline Form 1 ACN/water 980 200.4 50 suspension crystalline Form 1 ACN/water 950 50 0.6 5 suspension*n/a - no solid ACN/water 950 50 0.6 25 suspension crystalline Form 1ACN/water 950 50 0.6 50 suspension crystalline Form 1 ACN/water 850 1500.8 5 suspension* n/a - no solid ACN/water 850 150 0.8 25 suspensioncrystalline Form 1 ACN/water 850 150 0.8 50 suspension crystalline Form1 *re-dissolved on filtering

TABLE 9 XRPD results from the competitive slurry/water activityexperiments Volume Observation IPA/ Volume after ACN water WaterTemperature stirring for Solvent (μl) (μl) activity (° C.) 4 days XRPDresult IPA/water 980 20 0.2 5 suspension crystalline, Form 1 IPA/water980 20 0.2 25 suspension partially crystalline, Form 1 IPA/water 980 200.2 50 suspension partially crystalline, Form 1 IPA/water 960 40 0.4 5suspension crystalline, Form 1 IPA/water 960 40 0.4 25 suspensioncrystalline, Form 1 IPA/water 960 40 0.4 50 suspension crystalline, Form1 IPA/water 920 80 0.6 5 suspension crystalline, Form 1 IPA/water 920 800.6 25 suspension crystalline, Form 1 IPA/water 920 80 0.6 50 suspensioncrystalline, Form 1 IPA/water 840 160 0.8 5 suspension crystalline, Form1 IPA/water 840 160 0.8 25 suspension crystalline, Form 1 IPA/water 840160 0.8 50 suspension crystalline, Form 1 ACN/water 990 10 0.2 5suspension crystalline, Form 1 ACN/water 990 10 0.2 25 suspensioncrystalline, Form 1 ACN/water 990 10 0.2 50 suspension crystalline, Form1 ACN/water 980 20 0.4 5 suspension crystalline, Form 1 ACN/water 980 200.4 25 suspension crystalline, Form 1 ACN/water 980 20 0.4 50 suspensionpartially crystalline, Form 1 ACN/water 950 50 0.6 5 suspensioncrystalline, Form 1 ACN/water 950 50 0.6 25 suspension crystalline, Form1 ACN/water 950 50 0.6 50 solid on crystalline, Form 1 walls ACN/water850 150 0.8 5 suspension crystalline, Form 6 ACN/water 850 150 0.8 25suspension crystalline, Form 1 ACN/water 850 150 0.8 50 suspensioncrystalline, Form 3

Results show that Form 1 is more stable than Form 6 in all the IPA/waterexperiments regardless of the temperature or water activity. No evidenceof any remaining Form 6 material was seen in any of the IPA/waterexperiments after slurrying for 4 days.

Form 1 is also the most stable form in all the ACN/water experimentsexcept at 50° C. with a water activity of 0.8 (where Form 3 was made)and at 5° C. for a water activity of 0.8 (where Form 6 was made). Theexperiment that gave Form 3 material visually looked as though the solidhas completely dissolved at some point as the material was stuck to thewalls of the vial and there was no solid in the liquors. This mayexplain why the unexpected solvated form was produced in thisexperiment.

Therefore, the only conditions where Form 6 is more stable than Form 1is at 5° C. in ACN/water with a water activity of 0.8 (15 vol % water),i.e. at cold temperatures and very high water activities. Thus, if acrystallisation solvent with a low water activity is chosen, if thesolvent is wet, Form 6 may not be produced. The water activity (andtherefore the water content) depends on the solvent, alcohols have amuch lower water activity for a given water content than acetates,acetonitrile or acetone.

Example 6 Single Crystal Experiments

A full crystal structure of Form 2 was collected and solved. A summaryof structural data for Compound 1 Form 2 is provided in Table 10. Form 2crystallizes in the orthorhombic system, space group P2₁2₁2₁ with thefinal R1 [I>2σ(I)]=4.93%.

TABLE 10 Sample details and crystal data for Compound 1 Form 2 ParameterValue Crystallisation solvents DCM/acetone Crystallisation method Assupplied from column Empirical formula C₃₁H₂₆Cl₂N₈O₂ Formula weight613.50 Temperature 100(2) K Wavelength 1.54178 Å Crystal size 0.500 ×0.100 × 0.050 mm Crystal habit Colorless Prism Crystal systemOrthorhombic Space group P2₁2₁2₁ Unit cell dimensions a = 8.7289(3) Å α= 90° b = 13.2219(4) Å β = 90° c = 25.9554(7) Å γ = 90° Volume2995.59(15) Å³ Z 4 Density (calculated) 1.360 Mg/m³ Absorptioncoefficient 2.307 mm⁻¹ F(000) 1272

The asymmetric unit of Form 2 contains one fully ordered molecule ofCompound 1 and one molecule of dichloromethane See FIG. 21. Anisotropicatomic displacement ellipsoids for the non-hydrogen atoms are shown atthe 50% probability level. Hydrogen atoms are displayed with anarbitrarily small radius.

The absolute configuration of Compound 1 has been determined as depictedbelow with the Flack parameter=0.005(9). See Parsons, S and Flack, H.,Acta Cryst. 2004, A60, s61. The chiral center has an S configuration.

The DCM molecules sit in voids in the structure and are not bonded tothe API molecules. Without being bound to a particular theory, this mayexplain why some of the solvent can be removed on drying withoutdisrupting the crystal lattice and also why the same crystalline formcan be made from a range of solvents.

Table 11 provides the details on data collection and structurerefinement.

TABLE 11 Data collection and structure refinement for Compound 1 Form 2Parameter Value Diffractometer SuperNova, Dual, Cu at zero, AtlasRadiation source SuperNova (Cu) X-ray Source, CuKα Data collectionmethod omega scans Theta range for data 9.084 to 74.474° collectionIndex ranges −8 ≤ h ≤ 10, −16 ≤ k ≤ 16, −32 ≤ l ≤ 31 Reflectionscollected 31797 Independent reflections 6103 [R(int) = 0.0516] Coverageof independent 99.4% reflections Variation in check n/a reflectionsAbsorption correction Semi-empirical from equivalents Max. and min.transmission 1.00000 and 0.63134 Structure solution technique Directmethods Structure solution program SHELXTL (Sheldrick, 2013) Refinementtechnique Full-matrix least-squares on F² Refinement program SHELXTL(Sheldrick, 2013) Function minimized Σ w(F_(o) ² − F_(c) ²)²Data/restraints/parameters 6103/0/412 Goodness-of-fit on F² 1.039 Δ/σmax0.002 Final R indices 5464 data; I > 2σ(I) R1 = 0.0493, wR2 = 0.1335 alldata R1 = 0.0561, wR2 = 0.1428 Weighting scheme w = 1/[σ² (F_(o) ²) +(0.0956P)² + 0.7150P] where P = (F_(o) ² + 2F_(c) ²)/3 Absolutestructure parameter 0.005(9) Extinction coefficient n/a Largest diff.peak and hole 0.404 and −0.445 eÅ⁻³

Refinement summary: Ordered Non-H atoms, XYZ Freely refining OrderedNon-H atoms, U Anisotropic H atoms (on carbon), XYZ Idealized positionsriding on attached atoms H atoms (on carbon), U Appropriate multiple ofU(eq) for bonded atom H atoms (on heteroatoms), XYZ Freely refining Hatoms (on heteroatoms), U Isotropic Disordered atoms, OCC 2 part modelconstrained to unity Disordered atoms, XYZ freely refining Disorderedatoms, U anisotropic

Example 7 Cocrystal Screening

Materials

A diverse set of 28 pharmaceutically acceptable coformers was selectedbased on the potential bonding motifs of Compound 1. Compound 1 used inthis example is amorphous unless stated otherwise. The list ofconformers used for the cocrystal screen are: citric acid, L-malic acid,L-tartaric acid, fumaric acid, succinic acid, maleic acid, sorbic acid,ketoglutaric acid, salicylic acid, benzoic acid, 3-hydroxybenzoic acid,2,4-dihydroxybenzoic acid, 4-aminobenzoic acid, orotic acid, urea,nicotinic acid, isonicotinic acid, nicotinamide, isonicotinamide,saccharin, L-lactic acid, L-serine, L-proline, glycine, maltol,succinimide, sulfacetamide, and p-toluenesulfonic acid monohydrate.

Solvent Drop Grinding

A 1:1 stoichiometric mixture of Compound 1 and the coformer was weighedto produce a total of 50 mg in a 2 ml stainless steel grinding jar withone 7 mm grinding ball. The materials were wetted with solvent (10 μl ofnitromethane) and ground for 30 min at 30 Hz using a Retsch Mixer MillerMM300. Solids obtained after grinding were initially analyzed by XRPD.Sticky solids or liquids obtained after grinding were left to dry for 1day and any solids obtained were initially analyzed by XRPD. A controlexperiment with no conformer present was also performed following thesame procedure.

Dry Grinding

A 1:1 stoichiometric mixture of Compound 1 and the coformer was weighedto produce a total of 50 mg in a 2 ml stainless steel grinding jar withone 7 mm grinding ball. The materials were ground for 30 min at 30 Hzusing a Retsch Mixer Miller MM300. Solids obtained after grinding wereinitially analyzed by XRPD. A control experiment with no conformerpresent was also performed following the same procedure.

Sonication

A 1:1 stoichiometric mixture of Compound 1 (25 mg) and the coformer wasplaced in 2 ml HPLC vials and suspended in MeCN:water 1:1 (1 ml).Ultrasound was then applied in a cup horn at 50% power for 10 min withpulses of 30 s on and 1 min off. Solids obtained after sonication werefiltered and dried under suction for 10 min and the residues wereinitially analyzed by XRPD. Solutions obtained after sonication wereplaced at 5 and −20° C. for a day and any solids obtained were filteredand analyzed by XRPD. If no solids were obtained after cooling, sampleswere allowed to evaporate at RT and any solids obtained were analyzed byXRPD.

An initial control experiment with no coformer showed that Compound 1 isnot wettable in water alone. MeCN was added to produce a suspension,showing that MeCN:water 1:1 mixture produces a suspension.

The results of the experiments are provided below in Table 12.

TABLE 12 Cocrystal screen results Coformer Procedure Solvent IsolationObservation XRPD None Solvent drop Nitromethane Grinding Yellow solidForm 1 grinding Citric acid Solvent drop Nitromethane Grinding Yellowsticky Amorphous grinding solid L-Malic acid Solvent drop NitromethaneGrinding Yellow wet Form 1 grinding solid L-Tartaric acid Solvent dropNitromethane Grinding Yellow solid Form P1C3 grinding Fumaric acidSolvent drop Nitromethane Grinding Yellow solid Form 1 + grindingCoformer Succinic Acid Solvent drop Nitromethane Grinding Yellow wetAmorphous + grinding solid Coformer + Poor Form 1 Maleic Acid Solventdrop Nitromethane Grinding Yellow sticky Amorphous grinding solid Sorbicacid Solvent drop Nitromethane Grinding Yellow solid Form 1 + grindingCoformer Ketoglutaric acid Solvent drop Nitromethane Grinding Yellowsticky Amorphous + grinding solid 1 peak Salicylic acid Solvent dropNitromethane Grinding Yellow sticky Amorphous grinding solid Benzoicacid Solvent drop Nitromethane Grinding Yellow sticky Form 1 grindingsolid 3-Hydroxybenzoic Solvent drop Nitromethane Grinding Yellow sticky— acid grinding solid 2,4- Solvent drop Nitromethane Grinding Yellowsticky — Dihydroxybenzoic grinding solid acid 4-Aminobenzoic Solventdrop Nitromethane Grinding Yellow sticky Amorphous + acid grinding solidCoformer Orotic acid Solvent drop Nitromethane Grinding Yellow solidForm 1 + grinding Coformer Urea Solvent drop Nitromethane GrindingYellow solid Form 1 + grinding Coformer Nicotinic acid Solvent dropNitromethane Grinding Yellow solid Form 1 + grinding CoformerIsonicotinic acid Solvent drop Nitromethane Grinding Yellow solid Form1 + grinding Coformer Nicotinamide Solvent drop Nitromethane GrindingYellow solid Form 1 grinding Isonicotinamide Solvent drop NitromethaneGrinding Yellow solid Form 1 grinding Saccharin Solvent dropNitromethane Grinding Yellow solid Form 1 + grinding Coformer L-Lacticacid Solvent drop Nitromethane Grinding Yellow solid Similar to grindingForm 3 + Coformer L-Serine Solvent drop Nitromethane Grinding Yellowsolid Form 1 + grinding Coformer Nicotinic acid Solvent dropNitromethane Grinding Yellow solid Form 1 + grinding CoformerIsonicotinic acid Solvent drop Nitromethane Grinding Yellow solid Form1 + grinding Coformer Nicotinamide Solvent drop Nitromethane GrindingYellow solid Form 1 grinding Isonicotinamide Solvent drop NitromethaneGrinding Yellow solid Form 1 grinding Saccharin Solvent dropNitromethane Grinding Yellow solid Form 1 + grinding Coformer L-Lacticacid Solvent drop Nitromethane Grinding Yellow solid Similar to grindingForm 3 + Coformer L-Serine Solvent drop Nitromethane Grinding Yellowsolid Form 1 + grinding Coformer L-Proline Solvent drop NitromethaneGrinding Yellow solid Form 1 + grinding Coformer Glycine Solvent dropNitromethane Grinding Yellow solid Form 1 + grinding Coformer MaltolSolvent drop Nitromethane Grinding Yellow solid Form 1 + grindingCoformer Succinimide Solvent drop Nitromethane Grinding Yellow solidForm 1 grinding Sulfacetamide Solvent drop Nitromethane Grinding Yellowsolid Form 1 + grinding Coformer p-Toluenesulfonic Solvent dropNitromethane Grinding Yellow — acid-H₂O grinding liquid None Sonication1:1 Filtering Yellow solid Form 1 MeCN:Water Citric acid Sonication 1:1Filtering Yellow solid Similar to MeCN:Water Form 6 L-Malic acidSonication 1:1 Filtering Yellow solid Similar to MeCN:Water Form 6L-Tartaric acid Sonication 1:1 Filtering Yellow solid Similar toMeCN:Water Form 6 Fumaric acid Sonication + 1:1 Evaporation Yellow solidForm 1 + Evaporation MeCN:Water Coformer Succinic Acid Sonication 1:1Filtering Yellow solid Form 1 MeCN:Water Maleic Acid Sonication 1:1Filtering Yellow solid Form 1 MeCN:Water Sorbic acid Sonication 1:1Filtering Yellow solid Similar to MeCN:Water Form 6 Ketoglutaric acidSonication 1:1 Filtering Yellow solid Similar to MeCN:Water Form 6 + 5new peaks Salicylic acid Sonication 1:1 Filtering Yellow solid Form P1C9MeCN:Water Benzoic acid Sonication 1:1 Filtering Yellow solid Form 1MeCN:Water 3-Hydroxybenzoic Sonication 1:1 Filtering Yellow solid Form 1acid MeCN:Water 2,4- Sonication 1:1 Filtering Yellow solid Form 1Dihydroxybenzoic MeCN:Water acid 4-Aminobenzoic Sonication 1:1 FilteringYellow solid Form 1 acid MeCN:Water Orotic acid Sonication 1:1 FilteringYellow solid Similar to MeCN:Water Form 6 + Coformer Urea Sonication 1:1Filtering Yellow solid Similar to MeCN:Water Form 6 Nicotinic acidSonication 1:1 Filtering Yellow solid Similar to MeCN:Water Form 6Isonicotinic acid Sonication 1:1 Filtering Yellow solid Form 1 +MeCN:Water Coformer Nicotinamide Sonication 1:1 Filtering Yellow solidSimilar to MeCN:Water Form 6 Isonicotinamide Sonication 1:1 FilteringYellow solid Form 1 MeCN:Water Saccharin Sonication 1:1 Filtering Yellowsolid Form 1 MeCN:Water L-Lactic acid Sonication 1:1 Filtering Yellowsolid Form 1 MeCN:Water L-Serine Sonication 1:1 Filtering Yellow solidForm 1 MeCN:Water L-Proline Sonication 1:1 Filtering Yellow solid Form 1MeCN:Water Glycine Sonication 1:1 Filtering Yellow solid Form 1MeCN:Water Maltol Sonication 1:1 Filtering Yellow solid Similar toMeCN:Water Form 6 Succinimide Sonication 1:1 Filtering Yellow solidSimilar to MeCN:Water Form 6 Sulfacetamide Sonication 1:1 FilteringYellow solid Form 1 MeCN:Water p-Toluenesulfonic Sonication 1:1Filtering Yellowish Form 1 acid-H2O MeCN:Water solid None Dry grindingNone Grinding Yellow solid Amorphous L-Tartaric acid Dry grinding NoneGrinding Yellow solid Amorphous —: not performed,

TABLE 13 Preliminary characterization of the cocrystal screen sampleXRPD after 7 d XRPD High 40° C. 75% RH Coformer resolution ¹H-NMR TGADSC 25° C. 97% RH L-Tartaric Form Consistent 0.5% weight loss Smallbroad No change for acid P1C3 with between 30-100° C. endotherm (onsetboth. structure. 1.9% weight loss 33.5° C., 1.1 Eq. of between 100-160°C. peak 57.8° C., acid. 15.9% weight loss 5.27 J/g). Water peak between170-260° C., Endotherm (onset present. followed by 129.2° C., peakdegradation. 148.9° C., 78.48 J/g). L-Tartaric Form 6 Consistent 10.3%weight loss Large broad — acid with structure. between 30-100° C.endotherm (onset No peaks of No other weight 46.4° C., 274.0 J/g). theacid. loss until sample Small endotherm Water peak degrades (onset154.1° C., present. at ~240° C. 21.6 J/g). Small endotherm (onset 242.9°C., 4.9 J/g). Salicylic Form Consistent — Large broad — acid P1C9 withstructure. endotherm (onset 0.5 Eq. of acid 42.8° C., 149.6 J/g). Waterpeak Small endotherm present. (onset 120.3° C., 16.7 J/g). — = Notperformed.

Solvent-drop grinding experiments and dry grinding experiments resultedin mostly amorphous pattern, Form 1 or mixtures of Form 1 and Form 3with the coformer. Few experiments only produced sticky solids orliquids that were not suitable for XRPD analyses. One of the solventdrop grinding experiments (L-tartaric acid) resulted in a newcrystalline pattern (Form P1C3) that did not correspond to any of theknown crystalline forms of the Compound 1 or the coformer. XRPD, TGA andDSC characterizations of Form P1C3 are shown in FIGS. 22-23. GVSanalysis of Form P1C3 is shown in FIG. 28.

The sonication experiments performed resulted in mostly Form 1, Form 6or mixtures of Form 1 and Form 6 with the coformer. One of theexperiments (salicylic acid) resulted in a new crystalline pattern (FormP1C9) that did not correspond to any of the known crystalline forms ofthe Compound 1 or the coformer. XRPD, TGA and DSC characterizations ofForm P1C9 are shown in FIGS. 24-25.

Both samples displaying the new crystalline Forms P1C3 and P1C9 werecharacterized further. As the initial amount of Form 6 obtained in thepolymorphism screen was very limited, one of the samples that displayedForm 6 was also further characterized. The results are summarized inTable 13.

High resolution XRPD analysis of the potential cocrystal Form P1C3 wasconsistent with the low resolution XRPD, showing no presence of thestaring materials. ¹H-NMR showed that material was consistent with thestructure of Compound 1 with no evidence of degradation. The ¹H-NMR alsoshowed the presence of 1.1 eq. of the coformer and of water. Thermalanalyses of Form P1C3 show a DSC with a small broad endothermic eventbetween ˜RT and ˜100° C., which correlates with a 0.45% weight loss inthe TGA and a second large endothermic event (onset 129.2° C.), whichcorrelates with 1.9% weight loss in the TGA. The thermal data of FormP1C3 is consistent with a cocrystal solvate with 1 equivalent of water.Preliminary stability studies were carried out at 40° C./75% RH and 25°C./97% RH for Form P1C3 and no changes in the XRPD patterns wereobserved after a week.

High resolution XRPD analysis of the potential cocrystal Form P1C9 wasconsistent with the low resolution XRPD, showing no presence of thestaring materials. ¹H-NMR showed that material was consistent with thestructure of Compound 1 with no evidence of degradation. The ¹H-NMR alsoshowed the presence of 0.5 eq. of the coformer and of water. Thermalanalyses of Form P1C9 by DSC (no sample available for TGA analyses)showed clear differences to the starting materials, showing a broadendothermic event between ˜30 and 100° C., which indicate that thesecould be a solvated form.

High resolution XRPD analysis of the free base Form 6 was consistentwith the low resolution XRPD, showing no presence of the coformer or anyother forms of the free base. ¹H-NMR showed that material was consistentwith the structure of Compound 1 with no evidence of degradation and nopresence of the coformer. The ¹H-NMR also shows a large water peak. TheDSC analyses of Form 6 show a broad endothermic event between ˜30 and100° C., which indicate that this could be solvated form. The TGA ofForm 6 is also consistent with a solvate, showing a weight loss between˜30 and 100° C. that is consistent with 3 eq. of water.

In order to investigate the potential cocrystals for potentialdevelopment the forms obtained from L-tartaric (Form P1C3) and salicylicacids (Form P1C9) were selected for scale up and further studies.

Example 8 Scale Up Preparation of Cocrystals

Compound 1 used in this example is amorphous unless stated otherwise.

Slow Cooling

A 1:1 stoichiometric mixture of Compound 1 (40 or 400 mg) and thecoformer (L-tartaric acid or salicylic acid) was placed in 4 ml vialsand the appropriate solvent amounts were added to the vial. Samples wereplaced in a multireactor and were left at 40° C. (for DCM experiments)and 50° C. for other solvents. Samples were checked after 10 to 30 minof heating and if no dissolution was observed more solvent was addeduntil complete dissolution was reached. Dissolution was observed in allcases except for one experiment, which showed some coformer crystals insolution. For this experiment ethanol was added until completedissolution of the coformer was reached. The samples were then cooleddown to 5° C. at 0.1° C./min and 0.25° C./min. The resulting suspensionsor solutions were left at 5° C. for ca. 24 h. Any solids obtained werefiltered and analyzed by XRPD. Solutions obtained were placed at −20° C.for 2 h and any solids obtained were filtered and analyzed by XRPD. Ifno solids were obtained after cooling, samples were allowed to evaporateat RT and any solids obtained were analyzed by XRPD. The details and theresults of the scale-up experiments are summarized in tables below.

Sonication

A 1:1 stoichiometric mixture of Compound 1 at 25, 50, 250 or 500 mg andthe coformer (salicylic acid) was placed in a glass vial and suspendedin a MeCN-water or an ethanol-water mixture. Ultrasound was then appliedin a cup horn at 50% or 100% power for 10 min with pulses of 30 s on and1 min off. Solids obtained after sonication were allowed to stand at RTfor between 30 and 120 min and then filtered and dried under suction for10 min and the residues were analyzed by XRPD. The details and theresults of the experiments are summarized in tables below.

Characterization of Cocrystals

Certain scaled up cocrystal samples were characterized by: highresolution XRPD, VT-XRPD, DSC, TGA, ¹H-NMR, stability after 7 days at40° C./75% RH and at 25° C./97% RH, microscopy (PLM and SEM), GVS, KarlFischer, HPLC purity and kinetic solubility after 30 min in water and in0.1M HCl. See tables below for the summarized results.

Results

The results of the scale up and further characterization on selectedcocrystals are summarized in the following tables.

TABLE 14 Cocrystal scale up results (1/3) Solvent amount amount IsolatedCoformer Procedure (mg) Solvent (ml) after Final result XRPD L-TartaricSlow cooling 40 DMSO 0.150 Cooling to 5° C. Suspension in Similar toacid a yellow Form 2 and solution Form 4 L-Tartaric Slow cooling 40 2-0.150 Cooling to 5° C. Suspension in Form 1 + acid Methoxyethanol ayellow hints of Form 2 solution L-Tartaric Slow cooling 40 Nitromethane2.00 Cooling to 5° C. Suspension in Coformer acid (Bottle H02937) abrown solution L-Tartaric Slow cooling + 40 Dichlormethane- 5.00Evaporation Crystals in a Form 2 acid evaporation 40% ethanol yellowsolution L-Tartaric Slow cooling + 40 Nitromethane 2.00 EvaporationSuspension in Form P1C3 acid evaporation (Bottle H02937)- a brown 5%water solution Salicylic Slow cooling 40 DMSO 0.150 Cooling to 5° C.Suspension in Similar to acid a yellow Form 2 and solution Form 4Salicylic Slow cooling 40 2- 0.150 Cooling to 5° C. Suspension in Form1 + acid Methoxyethanol a yellow hints of Form 2 solution Salicylic Slowcooling + 40 Nitromethane 2.00 Evaporation Resin — acid evaporation(Bottle H02937) Salicylic Slow cooling + 40 Dichloromethane 3.00Evaporation Resin — acid evaporation Salicylic Slow cooling 40Nitromethane 2.00 Cooling to −20° C. Needles in a Similar to acid(Bottle H02937)- brown solution Form 6 5% water Salicylic Slow cooling +40 MeCN- 2.00 Evaporation Suspension in Form 1 acid evaporation 50%water a yellow solution — = Not performed

TABLE 15 Cocrystal scale up results (2/3) Time at RT Solvent Sonicationafter amount amount amplitude Result after sonication Coformer Procedure(mg) Solvent (ml) (%) sonication (min) XRPD Salicylic Sonication 25MeCN- 1.00 50 Suspension   30 min Form 1 acid 50% Water SalicylicSonication 50 MeCN- 1.00 50 Suspension   30 min Form P1C9 acid 50% WaterSalicylic Sonication 25 MeCN- 1.00 50 Suspension >120 min  Form P2C9 +acid 50% Water Form 1 Salicylic Sonication 50 MeCN- 1.00 50Suspension >120 min  Form P2C9 acid 50% Water Salicylic Sonication 50MeCN- 1.00 50 Suspension >60 min P1C9 + 1 acid 50% Water new peakSalicylic Sonication 50 MeCN- 1.00 50 Suspension >60 min Form P1C9 acid25% Water Salicylic Sonication 50 MeCN- 1.00 50 Suspension >60 min Form1 acid 5% Water Salicylic Sonication 50 Ethanol- 1.00 50 Suspension >60min Form 1 acid 50% Water Salicylic Sonication 50 Ethanol- 1.00 50Suspension >60 min Form 1 acid 25% Water Salicylic Sonication 50Ethanol- 1.00 50 Suspension >60 min Form 1 acid 5% Water SalicylicSonication 500 MeCN-50% 20.00 50 Suspension   60 min Form 1 acid waterSalicylic Sonication 500 MeCN-50% 10.00 50 Suspension   60 min Form 1acid water Salicylic Sonication 250 MeCN-50% 5.00 50 Suspension   60 minForm 1 acid water Salicylic Sonication 250 MeCN-50% 5.00 100Suspension >120 min  Form 1 acid water — = Not performed

TABLE 16 Cocrystal scale up results (3/3) Solvent Final amount amountIsolated amount Coformer Procedure (mg) Solvent (ml) after Final result(mg) XRPD L-Tartaric Slow 40 Nitromethane 2.00 Cooling to Suspension in 28.39 Form acid cooling + (Bottle 5° C. + 1 a yellow P1C9 evaporationH03010)- day solution 5% Water evaporation L-Tartaric Slow 400Nitromethane 20.00 Cooling to Brown-black — — acid cooling + (Bottle 5°C. + 4 resin evaporation H02937)- day 5% Water evaporation L-TartaricSlow 400 Nitromethane 20.00 Cooling to Suspension in 369.39 Form acidcooling + (Bottle 5° C. + 4 a yellow P1C9 evaporation H03010)- daysolution 5% Water evaporation — = Not performed

TABLE 17 Characterization of selected cocrystal samples XRPD and HPLCafter 7 d 40° C. High 75% RH Res. ¹H- 25° C. Form Coformer XRPD DSC TGANMR 97% RH KF Form L- Form 1st broad 1.2% Consistent XRPD: P1C3 tartaricP1C3 endotherm weight loss with no Acid onset corresponding structure.change ~30° C. to the 1.15 eq. for both. (11.95 J/g). first DSC of 2ndevent. coformer. endothermic 2.8% large weight loss peak correspondingonset at to the 120.94° C. second (65.86 J/g). DSC event. Form 6Salicylic — — — Consistent — Acid with structure. No coformer present.Form Salicylic — — 10.15% — — — P1C9 Acid weight loss from RT up to~100° C. Form Salicylic Form 1 large 5.64% Consistent — P2C9 Acid P2C9broad weight loss with endotherm corresponding structure. onset to the1.2 eq. of ~30° C. DSC coformer. and peak endothermic 0.48 eq. at 96.2°C. event. of (ΔH = 116.8 J/g). MeCN Form L- Form 1st broad 0.4%Consistent XRPD: 3.5% P1C3 tartaric P1C3 endotherm weight loss with noWater Acid onset corresponding the change ~30° C. to the structure. forboth. (6.907 J/g). first DSC 1.35 eq. HPLC: 2nd event. of shows aendothermic 2.8% coformer purity large weight loss 0.08 eq. change ofpeak corresponding of less than onset at to the MeNO₂. 3% for 124.64° C.second Other both. (67.93 J/g). DSC event. peaks observed. HPLC 30 minPurity Kinetic (%, solubility Form GVS Microscopy AUC) (pH) Form —Crystals of — 98.4% — P1C3 lath and plate-like morphology of typicallyless than 300 μm, most around 100 μm. Form 6 — Crystals of — — — —acicular morphology of typically less than 700 μm, most around 300 μm.Form — — — — — — P1C9 Form — — — — — — P2C9 Form 3.67% Lath Consistent88.4% Solubility Solubility P1C3 weight gain shaped with (sampledetermined determined from 0-90% crystals of microscopy 1) in in RH,cycle 2. sizes results 96.5% duplicate duplicate No between showing(sample as as hysteresis ~5 to 750 μm lath 2) 0.04 0.02 observed. long.shaped and and XRPD after crystals 0.05 mg/ml 0.02 mg/ml analysis with(pH (pH shows no smooth 2.9, 1.3) change. flat 3.0) surfaces and sharpedges. — = not performed, * = free base Compound 1 solubility determinedas 0.01 and 0.01 mg/ml (pH 7.1 and 6.8), ** = free base Compound 1solubility determined as 0.01 and 0.01 mg/ml (pH 1.5 and 1.3).Scale Up of L-tartaric Acid Cocrystal Form P1C3

Preliminary slow cooling experiments with L-tartaric acid showed thatwhile four of the solvents used resulted in crystallization of the freebase or of the coformer, the experiment performed using MeNO₂-5% waterresulted in a solution that after 1 day evaporation produced thecocrystal Form P1C3.

Following the small scale investigations further slow cooling andevaporation experiments were carried out at 500 mg scale using MeNO₂-5%water in order to produce the L-tartaric acid cocrystal Form P1C9. Allthe preliminary small scale experiments performed with MeNO₂ showed abrown solution which were related to some potential contamination ordegradation of the MeNO₂.

Characterization of the sample obtained from L-tartaric acid showed thatthe XRPD was consistent with the Form P1C3 previously obtained in thecocrystal screen with no evidence of coformer or free base formspresent. Thermal analysis showed a DSC with a first small broadendothermic event between ˜RT and ˜100° C., which correlates with a0.45% weight loss in the TGA. A second large endothermic event in theDSC with onset at 124.6° C. correlates with 2.8% weight loss in the TGA.These two thermal events indicate some initial residual solvent lossbefore 100° C., followed by solvent loss corresponding to c.a. 1equivalent of water (calculated 2.5% water for a 1:1:1 Compound 1:L-tartaric acid: water cocrystal). The Karl Fischer analyses showed 3.5%water content, which is also consistent with a cocrystal hydrate.VT-XRPD showed no change until 140° C., where the sample shows anamorphous pattern, which does not change after cooling the sample to 30°C. ¹H-NMR shows that the material is consistent with the structure ofCompound 1 and it also shows peaks corresponding to the coformer thatintegrate as 1.35 eq. (note that there is only one diagnostic peak ofthe coformer). GVS analyses show that the material is moderatelyhygroscopic with 3.67% weight gain from 0-90% RH. No hysteresis isobserved and the XRPD post GVS shows no change Analysis by PLM and SEMshow only one type of crystal habit with the same lath morphology and ofsizes ranging between 5 and 750 μm. Stability under differenttemperature and RH conditions showed no significant change in the purityor in the XRPD after a week at 25° C./96% RH and at 40° C./75% RH.Kinetic solubility determined after 30 min in water and 0.1M HCl showedvalues very similar to the free base solubility for the 0.1M HClexperiments, but a slightly higher solubility values were obtained forthe water experiments. The XRPD of the solubility residues showed nochange with respect to the staring materials.

Scale Up of Salicylic Acid Cocrystal Forms P1C9 and P2C9

Preliminary slow cooling experiments with salicylic acid only producedcrystallization of polymorphic forms of the free base or the formationof gums.

Preliminary sonication experiments were performed following the sameprocedure that initially produced the salicylic acid cocrystal FormP1C9, but the materials were allowed to settle for longer times beforefiltering to potentially increase the yield. However, these experimentsshowed that if the sample was allowed to settle for more than 120 min anew pattern described as P2C9 that did not correspond to the known freebase forms or to the coformer was obtained. Also the experiments showedthat even when the sample was allowed to settle only for 30 min thecocrystal Form P1C9 was not consistently obtained, producing thecocrystal Form P1C9 for the 50 mg scale experiment and free base Form 1for the 25 mg scale.

Characterization of the sample obtained from salicylic acid showed thatthe XRPD was consistent with the Form P1C9. The TGA analyses showed a10.15% weight loss before 100° C. that correlates with the firstendothermic event observed in the DSC of Form P1C9. This together withthe previous data obtained for Form P1C9 is consistent with a cocrystalhydrate with ˜4 eq. of water.

Characterization of the sample obtained from salicylic acid showed a newpotential cocrystal Form P2C9. XRPD, TGA and DSC characterizations ofForm P2C9 are shown in FIGS. 26-27. Thermal analysis showed a DSC with alarge broad endothermic event between RT and ˜110° C., which correlateswith a 5.64% weight loss in the TGA. ¹H-NMR shows that the material isconsistent with the structure of Compound 1 and it also shows peaks thatintegrate as 1.2 eq. of the coformer, as well as the MeCN peak thatintegrates as 0.48 eq. of MeCN. Thermal data together with the ¹H-NMRdata indicate that P2C9 is a salicylic acid cocrystal solvate with MeCN(calculated 5.6% MeCN for a 1:1:1 Compound 1: salicylic acid: MeCNcocrystal).

A cocrystal screen was carried out on Compound 1 using 28 coformers.Sonication and solvent-drop grinding methodologies were used for thecocrystal screen.

Two different cocrystals were identified in the screen from 2 differentcoformers: cocrystal Form P1C3 obtained from L-tartaric and cocrystalForm P1C9 obtained from salicylic acid. These potential cocrystalsshowed diffractograms that did not correspond to any of the knowncrystalline forms of the Compound 1 or the coformers. Further ¹H-NMRcharacterization on cocrystal Forms P1C3 and P1C9 confirmed the presenceof the coformer, with 1.1 and 0.5 eq. of acid respectively. Thermal DSCand TGA analyses also indicated cocrystal formation in both casesshowing clear differences with respect to the starting materials.

Both cocrystal Forms P1C3 and P1C9 obtained from L-tartaric andsalicylic acids were selected for scale up and further investigations.

The L-tartaric acid cocrystal Form P1C3 was successfully scaled up to c.a. 500 mg from slow cooling and evaporation and fully characterized,which confirmed that Form P1C3 is a 1:1:1 Compound 1:L-tartaric acid:water cocrystal hydrate. Kinetic solubility studies on the cocrystalForm P1C3 showed a slight improve in solubility with respect to the freebase. Attempts to produce the salicylic acid cocrystal Form P1C9 fromslow cooling and evaporation experiments were unsuccessful, even in asmall scale. Preliminary smaller scale attempts to produce Form P1C9from sonication resulted in the formation of Form P1C9 in someinstances, however some of these experiments also produced a newpotential cocrystal Form P2C9. Further ¹H-NMR and thermalcharacterization of this new Form P2C9 indicated that the material waspotentially an acetonitrile cocrystal solvate, so no further scale upwas attempted. Attempts to produce Form P1C9 on a 500 mg scale fromsonication experiments only produced the free base Form 1. Furthercharacterization on the available Form P1C9 indicated that the materialis likely to be a 1:0.5:4 Compound 1:salicylic acid: water cocrystalhydrate.

Example 9 Solid Form Characterization

X-Ray Powder Diffraction (XRPD)

Bruker AXS C2 GADDS: X-Ray Powder Diffraction patterns were collected ona Bruker AXS C2 GADDS diffractometer using Cu Kα radiation (40 kV, 40mA), automated XYZ stage, laser video microscope for auto-samplepositioning and a HiStar 2-dimensional area detector. X-ray opticsconsists of a single Göbel multilayer mirror coupled with a pinholecollimator of 0.3 mm. A weekly performance check is carried out using acertified standard NIST 1976 Corundum (flat plate). The beam divergence,i.e. the effective size of the X-ray beam on the sample, wasapproximately 4 mm. A θ-θ continuous scan mode was employed with asample-detector distance of 20 cm which gives an effective 2θrange of3.2°-29.7°. Typically the sample was exposed to the X-ray beam for 120seconds. The software used for data collection was GADDS for XP/20004.1.43 and the data were analyzed and presented using Diffrac Plus EVAv15.0.0.0. Samples run under ambient conditions were prepared as flatplate specimens using powder as received without grinding. Approximately1-2 mg of the sample was lightly pressed on a glass slide to obtain aflat surface. Samples run under non-ambient conditions were mounted on asilicon wafer with heat-conducting compound. The sample was then heatedto the appropriate temperature at 10° C./min and subsequently heldisothermally for 1 minute before data collection was initiated.

Bruker AXS D8 Advance:

X-Ray Powder Diffraction patterns were collected on a Bruker D8diffractometer using Cu Kα radiation (40 kV, 40 mA), θ-2θgoniometer, anddivergence of V4 and receiving slits, a Ge monochromator and a Lynxeyedetector. The instrument is performance checked using a certifiedCorundum standard (NIST 1976). The software used for data collection wasDiffrac Plus XRD Commander v2.6.1 and the data were analyzed andpresented using Diffrac Plus EVA v15.0.0.0.

Single Crystal X-Ray Diffraction (SCXRD)

Data were collected on an Oxford Diffraction Supernova Dual Source, Cuat Zero, Atlas CCD diffractometer equipped with an Oxford CryosystemsCobra cooling device. The data was collected using CuKα radiation.Structures were typically solved using either the SHELXS or SHELXDprograms and refined with the SHELXL program as part of the Bruker AXSSHELXTL suite (V6.10). Unless otherwise stated, hydrogen atoms attachedto carbon were placed geometrically and allowed to refine with a ridingisotropic displacement parameter. Hydrogen atoms attached to aheteroatom were located in a difference Fourier synthesis and wereallowed to refine freely with an isotropic displacement parameter.

Nuclear Magnetic Resonance (NMR)

NMR spectra were collected on a Bruker 400 MHz instrument equipped withan auto-sampler and controlled by a DRX400 console. Automatedexperiments were acquired using ICON-NMR v4.0.7 running with Topspinv1.3 using the standard Bruker loaded experiments. For non-routinespectroscopy, data were acquired through the use of Topspin alone.Samples were prepared in DMSO-d₆, unless otherwise stated. Off-lineanalysis was carried out using ACD Spectrus Processor 2012.

Differential Scanning Calorimetry (DSC)

TA Instruments Q2000: DSC data were collected on a TA Instruments Q2000equipped with a 50 position auto-sampler. The calibration for thermalcapacity was carried out using sapphire and the calibration for energyand temperature was carried out using certified indium. Typically 0.5-3mg of each sample, in a pin-holed aluminium pan, was heated at 10°C./min from 25° C. to 375° C. A purge of dry nitrogen at 50 ml/min wasmaintained over the sample. Modulated temperature DSC was carried outusing an underlying heating rate of 2° C./min and temperature modulationparameters of ±0.64° C. (amplitude) every 60 seconds (period). Theinstrument control software was Advantage for Q Series v2.8.0.394 andThermal Advantage v5.5.3 and the data were analyzed using UniversalAnalysis v4.5A.

Mettler DSC 823e: DSC data were collected on a Mettler DSC 823E equippedwith a 34 position auto-sampler. The instrument was calibrated forenergy and temperature using certified indium. Typically 0.5-3 mg ofeach sample, in a pin-holed aluminium pan, was heated at 10° C./min from25° C. to 375° C. A nitrogen purge at 50 ml/min was maintained over thesample. The instrument control and data analysis software was STARev12.1.

Thermo-Gravimetric Analysis (TGA)

TA Instruments Q500: TGA data were collected on a TA Instruments Q500TGA, equipped with a 16 position auto-sampler. The instrument wastemperature calibrated using certified Alumel and Nickel. Typically 5-10mg of each sample was loaded onto a pre-tared aluminium DSC pan andheated at 10° C./min from ambient temperature to 450° C. A nitrogenpurge at 60 ml/min was maintained over the sample. The instrumentcontrol software was Advantage for Q Series v2.5.0.256 and ThermalAdvantage v5.5.3 and the data were analyzed using Universal Analysisv4.5A.

Mettler TGA/SDTA 851e:

TGA data were collected on a Mettler TGA/SDTA 851e equipped with a 34position auto-sampler. The instrument was temperature calibrated usingcertified indium. Typically 5-30 mg of each sample was loaded onto apre-weighed aluminium crucible and was heated at 10° C./min from ambienttemperature to 450° C. A nitrogen purge at 50 ml/min was maintained overthe sample. The instrument control and data analysis software was STARev12.1.

Water Determination by Karl Fischer Titration (KF)

The water content of each sample was measured on a Metrohm 874 OvenSample Processor at 150° C. with 851 Titrano Coulometer using HydranalCoulomat AG oven reagent and nitrogen purge. Weighed solid samples wereintroduced into a sealed sample vial. Approx 10 mg of sample was usedper titration and duplicate determinations were made. Data collectionand analysis using Tiamo v2.2.

Gravimetric Vapour Sorption (GVS)

Sorption isotherms were obtained using a SMS DVS Intrinsic moisturesorption analyser, controlled by DVS Intrinsic Control software v1.0.1.2(or v 1.0.1.3). The sample temperature was maintained at 25° C. by theinstrument controls. The humidity was controlled by mixing streams ofdry and wet nitrogen, with a total flow rate of 200 ml/min The relativehumidity was measured by a calibrated Rotronic probe (dynamic range of1.0-100% RH), located near the sample. The weight change, (massrelaxation) of the sample as a function of % RH was constantly monitoredby the microbalance (accuracy ±0.005 mg). Typically 5-20 mg of samplewas placed in a tared mesh stainless steel basket under ambientconditions. The sample was loaded and unloaded at 40% RH and 25° C.(typical room conditions). A moisture sorption isotherm was performed asoutlined below (2 scans giving 1 complete cycle). The standard isothermwas performed at 25° C. at 10% RH intervals over a 0-90% RH range. Dataanalysis was carried out using Microsoft Excel using DVS Analysis Suitev6.2 (or 6.1 or 6.0).

Example 10 Micronized Compound 1 Form 1

Micronized Compound 1 Form 1 was prepared using a Fluid EnergyJet-O-Mizer 00 Jet Mill with a 2″ loop. Approximately 9 g of Compound 1Form 1 was passed through the jet-mill twice with parameters outlinedbelow.

Supply Grinding Pusher Air Pressure Nozzle Pressure Nozzle PressureFeeder (psi) (psi) (psi) Speed 114 90 80 3

Approximately 7.3 g of micronized Compound 1 Form 1 was collected (81%yield). The particle size distribution after each pass is measured bythe wet dispersion method (using water as dispersant) on a MalvernMastersizer 2000 and the results are provided in the table below.

Particle Size (μm) Sample D₁₀ D₅₀ D₉₀ Unmilled* 2.7 12.0 40.0 1^(st)Pass 0.3 2.1 6.4 2^(nd) Pass 0.2 1.4 5.3

Example 11 Synthesis and Recrystallization of Compound 1

In a 50 L jacketed reactor equipped with an overhead stirrer,thermocouple and condenser was placed Compound C (1000 g, 2714 mmol) andCompound D (747 g, 2714 mmol) in 28.7 L (28.7 vol relative to CompoundC, 20 vol relative to the product Compound 1) 4:1 DCM:EtOH. Thesuspension was stirred 36° C. for 21 h and the reaction conversion wasmonitored by HPLC.

After the reaction was stopped, 1M KOH (152 g, 2714 mmol) in EtOH wascharged to the reaction. The mixture was stirred at 36° C. for 2 h, andthen the temperature was adjusted to 20° C. over 1 h. 7.2 L DI water wascharged and stirred for 15 min. The phases were allowed to separate. Thebottom organic layer was drained to 3 carboys. The aqueous layer wasdrained and saved. The reactor was cleaned with DI water and EtOH. Theorganic layer and another 7.2 L DI water were charged back to thereactor. The carboys were rinsed with 3.6 L 4:1 DCM:EtOH (2.5 volrelative to product Compound 1) and the rinse was combined with the restof the organic phase. The mixture was stirred for 15 min. The bottomorganic layer was drained to the carboys and kept overnight at rt. Theaqueous layer was drained and saved. The reactor was disassembledcompletely and cleaned with acetone, DI water and EtOH and purgedovernight with nitrogen.

The distillation setup was assembled onto the reactor. The organic layerwas pumped through a capsule filter (polycap, PTFE, 1 μm, cat#2603T)back into the clean reactor using a pump (KNF lab liquiport). The flaskwas rinsed with 3.6 L 4:1 DCM:EtOH (2.5 vol relative to productCompound 1) and the rinse was pumped through the capsule filter to becombined with the rest of the organic layer.

Distillation was started with stirring (controller set to 30). Jackettemperature was to 45° C. during the distillation. Pot temperature wasabout 23-27° C. during the distillation. Highest vacuum was about 400torr. The volume of solution was reduced to about 14.4 L (10 vol). Thesolution remained clear at the end of the distillation. Some solidformed on the wall of the reactor above the solution. The solution wasstirred (controller set at 20) overnight. Jacket temperature was set to25° C.

28.8 L (20 vol) EtOH was charged. The distillation was restarted. Jackettemperature was set to 55° C. Pot temperature was about 44-47° C.Highest vacuum was about 250 torr. The volume of solution was reduced toabout 1.5 L (15 vol). Solid formed during the distillation.

A mixture of 750 ml (7.5 vol) EtOH and 750 ml (7.5 vol) DI water wascharged to the suspension. The internal temperature was adjusted to 60°C. The solids redissolved as the temperature increased. No solid wasleft when temperature reached 60° C. 1 g of Compound 1 (1 wt %) wascharged as seed. Not all the seeds dissolved. The mixture was stirred at60° C. for a total of 6 h, cooled to 20° C. over 10 h, and then stirredat 20° C. for 6 h. The solid was collected by filtration through aBuchner funnel equipped with a filter paper. The filter cake was washedwith 2×500 ml 1:4 EtOH:water (2×5 vol). The solid was dried in theBuchner funnel with house vac for 2 d. The cake was then dried in avacuum oven at 40° C. with a nitrogen bleed overnight for 18 h. Theproduct Compound 1 was isolated as an off white solid. HPLC (˜0.3 mg/mlin 1:1 acetonitrile:water), NMR and DSC of the isolated solid wasobtained. The DSC indicates the solid is Form 1. NMR indicates very lowlevel of residual EtOH is present in the solid.

Compared to Method 3 of Example 1, comparable chemical and chiral puritywere obtained. Reaction was telescoped to polish filtration andrecrystallization. Isolation of crude Compound 1 was eliminated. Lessnumber of solvents were used. The process was slower, and the productcontained residual ethanol.

In order to reduce the residual ethanol content in the product, variousmodifications to the recrystallization conditions were investigated. Theresults are listed in the table below.

Ethanol DCM Ethanol content Purity content content (NMR mole YieldConditions (area %) (GC) (GC) ratio) (%) Chase with EtOH 99.5 445 ppm10590 ppm  0.10 90 Chase with 80% EtOH/H₂O 99.6 — — 0.06 80 Chase with60% EtOH/H₂O 99.6 ND 9623 ppm 0.07 82 Chase with 40% EtOH/H₂O 99.5 252ppm 25315 ppm  0.18 85 Chase with 80% EtOH/H₂O 98.6* ND 6926 ppm 0.05 77Chase with EtOH to 15 vol, 99.6 196 ppm 6981 ppm 0.05 80 add 5 vol H₂O,then seed Chase with EtOH to 15 vol, 99.6** — — 0.03 75 add 5 vol H₂O,then seed, 4 heat/cool cycles (3 h at 60° C., cool to rt in 2 h) Chasewith EtOH to 15 vol, 99.8 — — 0.05 78 add 5 vol H₂O, then seed, 3heat/cool cycles (3 h at 50° C., cool to rt in 2 h) Chase with EtOH to15 vol, 99.8 — — 0.03 72 add 5 vol H₂O, then seed, 2x final volume*containing Compound D. Compound D can be removed by quenching with KOHin ethanol. **containing 0.11% area of impurity

FIG. 29 shows Compound 1 solubility in ethanol/water and correspondingethanol content in isolated Compound 1. The results show that usingsolvent mixture with higher Compound 1 solubility during chase resultslower residual ethanol content in isolated Compound 1. Prolonged dryingdid not affect residual ethanol content (data not shown).

A crystal structure of Form 1 was collected and solved. FIG. 30 showsthe crystal structure of Form 1 of Compound 1 obtained by heating 13.5mg/ml Compound 1 in 80% ethanol/water to 60° C. then cool to rt. Thereis no ethanol in unit cell. A summary of structural data for Compound 1Form 1 is provided in the table below. Form 1 crystallizes in theorthorhombic system, space group P2₁2₁2₁.

Parameter Value Formula C₂₅H₃₁NO₆ D_(calc.)/g cm⁻³ 1.279 μ/mm⁻¹ 0.744Formula Weight 441.51 Color colorless Shape chunk Max Size/mm 0.30 MidSize/mm 0.15 Min Size/mm 0.15 T/K 173(2) Crystal System orthorhombicFlack Parameter −0.03(8) Hooft Parameter 0.01(8) Space Group P212121 a/Å11.14000(10) b/Å 12.75790(10) c/Å 16.1306(2) α/° 90 β/° 90 γ/° 90 V/Å³2292.53(4) Z 4 Z′ 1 Θ_(min)/° 4.418 Θ_(max)/° 72.157 Measured Refl.14293 Independent Refl. 4454 Reflections Used 4179 R_(int) 0.0301Parameters 294 Restraints 0 Largest Peak 0.172 Deepest Hole −0.237 GooF1.041 wR₂ (all data) 0.0928 wR₂ 0.0904 R₁ (all data) 0.0393 R₁ 0.0364

Example 12 Screening Reaction Conditions for Preparation of Compound Aby Amide Coupling of Compound C and Compound G

Different coupling reagents were investigated for the preparation ofCompound 1 through direct amide coupling of Compound C and Compound G.These reagents include: DECP, HDMA, HBTU, COMU, PyBOP, HATU, DEPC, DPPA,and BOPC1.

EDCI coupling reactions with various activators were also investigatedusing the following condition: EDCI (1.15 eq), Et₃N (1.1 eq), 4:1MeCN/H₂O (20 vol), 21° C. The results are listed in the Table below.

EDCI + Activator HPLC purity Activator Equiv (time) HOBt 1.1 94% (16 h)HOBt 0.2 74% (91 h) HBTriazinone 1.1 75% Oxyma 1.1 58% Oxyma 1.5 56%Oxyma 1.1 35% NHS 1.1 33% Oxyma 2.0 31% No activator 0 20% K-Oxyma 1.116%

The following phosphorus-based coupling reagents were also investigated:DEPBT (59.5% IPC purity after 51 hours), BrOP (30.4% IPC purity after 51hours), (EtO)₂P(O)—Cl (3-10% IPC purity), (EtO)₂P(O)-Oxyma (62.2% IPCpurity, 96% purity after isolation), and T3P (72.0% IPC purity after 48hours, base: pyridine, Temp: 40° C., solvent: MeCN (20 vol)). A scale uprun was carried out with T3P (3.0 eq.) and pyridine (5.1 eq.) at 40° C.for 2 days (63.8% IPC purity).

The following mixed anhydride coupling reagents were also investigated:pivaloyl chloride (15.8% IPC purity), and iso-butyl chloroformate (34.9%IPC purity in MeCN, 14.0% IPC purity in 4:1 MeCN:H₂O).

The following triazine-based coupling reagents were also investigated:CDMT and DMTMM.

An example reaction of DMTMM coupling was carried out as the follows. To1.15 eq DMTMM, 1.1 eq Et₃N (or DIPEA), 1.05 eq Compound G was charged4:1 MeCN:H₂O. The reaction mixture was stirred and activated overnight.1.0 eq Compound C was then added in one bolus as a solid. The reactiongave 84% HPLC purity after 21 hours.

The reaction mixture was warmed to 40° C. Water (10.5 vol) was addedover 2 h. The mixture was cooled to 21° C. over 2 h, then filtered,rinsed with 2:1 H₂O:MeCN (2×5 vol), and dried to provide granuleproduct. Crude yield (79.6-84.4%), purity (97.2-97.5%).

The crude product was further purified by polish filtration in 4:1DCM:EtOH, followed up recrystallization from 3:1 EtOH:H₂O (30 vol).Overall yield (56-62%), final purity (99.5-99.6%).

Further screenings for DMTMM coupling conditions were also carried outfor solvent (3:1 EtOH:H₂O, 14.4:4.8:1 EtOH:H₂O:DCM, 4:1 acetone:H₂O, 4:1DCM/MeOH, 4:1 DCM/EtOH, and 100% DCM); reaction volumes (5, 7.5, 10, 15,and 20); reaction temperature (5, 20, and 40° C.); base (Et₃N, iPr₂NEt,pyridine, NMM, DBU, NaOH, and DMAP); base equivalents (3.0, 1.1, and0.2); DMTMM equivalents (1.15, 1.10, 1.05, and 1.00); Compound Caddition (as a solid one bolus, and in solution over time); and additionof water after reaction reaches completion.

Example 13 Controlled Crystallization and Recrystallization of Compound1

Controlled crystallization and recrystallization of Compound 1 from asolvent of a mixture of acetonitrile and water were studied.

Controlled Crystallization Procedure

A reaction mixture was produced following Method 3 of Example 1 (afterquench of the reaction), consisting of Compound 1 at ˜124 mg/mlconcentration (˜8.06 L/kg volume), in a 90/10 v/v acetonitrile/watermixture.

An appropriate amount of acetonitrile/water were added to bring thesolution to 80/20 v/v acetonitrile/water at 20 L/kg of Compound 1. Theslurry was agitated and heated to 75° C. until all the Compound 1 wasdissolved. The clear solution was polished filtered with a Zapcap filter(0.45 μm). The solution was put back into the reactor and cool down to65° C. Form 1 of Compound 1 was used to seed with 10 wt % of theestimated amount of Compound 1 in the slurry (check that the seeds didnot dissolve). The mixture was cooled down to 22° C. over 16 hours(solubility was brought from ˜50 mg/ml to ˜18 mg/ml at a rate of 2mg/ml/hr) with agitation, and held under agitation for 2 hours aftercompletion of cooling step.

20 L/kg of water was added to bring the solution to 40/60 v/vacetonitrile/water (total volume of 40 L/kg of Compound 1) over 4 hours(solubility is brought from ˜18 mg/ml to ˜3 mg/ml) with a syringe pump.The mixture was held under agitation at 22° C. for 2 hrs after the endof water addition.

The slurry was filtered with a Buchner funnel, the cake was washed twicewith a 40/60 v/v acetonitrile/water mixture. The Buchner funnel wasplaced into oven at 60° C. under vacuum with nitrogen bleed to dryresidual solvents and provide Form 1 of Compound 1. In one exemplaryrun, the yield was 75%, and the purity was 99.7 area %.

The controlled crystallization procedure was also carried out on areaction mixture produced following the DMTMM coupling in Example 12. Inone exemplary run, the yield was about 90%, and the purity was 97.6 area%.

Controlled Recrystallization Procedure

Compound 1 was synthesized following Method 3 of Example 1 (afterisolation from acetonitrile/water).

20 L/kg of 80/20 v/v acetonitrile/water was added to Compound 1 solidcharged into a reactor. The slurry was agitated and heated to 75° C.until all Compound 1 was dissolved. The clear solution was polishfiltered with a Zapcap filter (0.45 μm). The solution was put back intothe reactor and cooled down to 65° C. Form 1 of Compound 1 was used toseed with 10 wt % of the estimated amount of Compound 1 in the slurry(check that the seeds did not dissolve). The mixture was cooled down to22° C. over 16 hours (solubility was brought from ˜50 mg/ml to ˜18 mg/mlat a rate of 2 mg/ml/hr) with agitation, and held under agitation for 2hours after completion of cooling step.

20 L/kg of water was added to bring the solution to 40/60 v/vacetonitrile/water (total volume of 40 L/kg of Compound 1) over 4 hrs(solubility was brought from ˜18 mg/ml to ˜3 mg/ml) with a syringe pump.The mixture was held under agitation at 22° C. for 2 hrs after the endof water addition.

The slurry was filtered with a Buchner funnel. The cake was washed twicewith a 40/60 v/v acetonitrile/water mixture. The Buchner funnel wasplaced into oven at 60° C. under vacuum with nitrogen bleed to dryresidual solvents and provide Form 1 of Compound 1. In one exemplaryrun, the purity was 99.7 area %.

The controlled crystallization procedure was also carried out on areaction mixture produced following the DMTMM coupling in Example 12. Inone exemplary run, the yield was about 75%, and the purity was 99.4 area%.

Example 14 Preparation of Amorphous Compound 1

Amorphous compound 1 can be prepared by dissolving a crystalline solidor a mixture of crystalline solids and amorphous sample in one or moresolvents and preforming lyophilisation on the solution. Amorphouscompound 1 can also be prepared by preforming lyophilisation on asolution of the material obtained after purification through columnchromatography. Other methods of preparing amorphous compound includespray drying.

Examples of spray solutions are described below with their respectivecompositions:

Spray Spray Spray Solution #2 Solution #3 Solution #1 (2:1 Compound (2:1Compound Component (100% Compound 1) 1:PVP/VA 64) 1:HPMC-AS) Compound 18.14 g 6.33 g 6.33 g Polymer — 3.15 g 3.15 g DCM 48 mL 56 mL 56 mL(63.84 g) (74.48 g) (74.48 g) MeOH 12 mL 14 mL 14 mL (9.50 g) (11.09 g)(11.09 g) % Solids 10% 10% 10% Content (w/w)

The DCM and MeOH were added to a glass jar and mixed. If needed, thepolymer was added to the solvent matrix while stirring and the solutionwas allowed to stir until all solids dissolved. Compound 1 was thenadded to the solution while stirring. All three spray solutions weremixed for an hour prior to spray drying. Spray drying was done using theBuchi B-290 mini lab-scale spray dryer and the spray parameters areprovided below:

Aspirator Inlet Outlet Nozzle Pump Spray Rate Temp Temp. Pressure SpeedRate (%) (° C.) (° C.) (psi) (%) (g/min)* 100 95 63-65 30 20 10 *pumpwas calibrated using 80:20 DCM:MeOH and might not be representative ofthe spray rate of the spray solutions with polymer due to the differencein viscosity

The spray dried products were dried in a vacuum oven at 25° C. for ˜22hours. The post-drying products were analyzed on TGA and DSC and theresults are provided in the table below.

Thermograms for Compound 1 Spray Dried SDDs−t=0

Sample TGA Comments Spray Dried FIG. 31 Expected T_(g): 147° C. Compound1 Observed T_(g): 146° C. 2:1 Compound FIG. 32 Predicted T_(g): 132° C.1:PVP/VA 64 Observed T_(g): 136° C. 2:1 Compound FIG. 33 PredictedT_(g): 143° C. 1:HPMC-AS Observed T_(g): 133° C.

Example 15 Formulation of Compound 1

Compound 1 was spray dried according to the methods described herein.The sample was blended with excipients, which was then encapsulated,polished and packaged. Below is a table of the formulation for 5 mgcapsule and 30 mg capsule.

5 mg capsule 30 mg capsule Component % w/w mg % w/w mg Compound 1 spraydried 1.92% 5.00 9.38% 30.00 (Neat Amorphous API) Starch 1500 46.29% 120.35 42.56%  136.20 (Pre-gelatinized Starch) Parteck M100 (Mannitol)46.29%  120.35 42.56%  136.20 Ac-Di-Sol 5.00% 13.00 5.00% 16.00(Croscarmellose Sodium) Magnesium Stearate 0.50% 1.30 0.50% 1.60 TOTAL 100% 260.00  100% 320.00 HPMC Capsule Shell Size 2, Swedish Orange Size1, White

Example 16 Screening Reaction Conditions for Preparation of Compound C

Metal and Pd Source Screen

Different metal sources with XPhos and alternative bases wereinvestigated. Pd catalysts generally work better than Ni catalyst(NiCl₂(PPh₃)₂). Cu catalyst (Cup was also explored as a co-catalyst tothe Pd catalyst or Ni catalyst.

Xphos-Pd-G3 and Pd₂(dba)₃/Xphos (1:1 L:M) worked as well as controlreaction but without observed delay. Pd₂(dba)₃: Xphos (1:1 L:M) workedwell with K₂CO₃ and K₃PO₄. PdCl₂(MeCN)₂: XPhos (2:1 L:M) worked wellwith K₂CO₃ and K₃PO₄. Pd(OAc)₂: XPhos worked better with 2:1 L:M ratiothan 1:1 ratio. Pd(PPh₃)₄ and PdCl₂(PPh₃)₂ also worked. The relativerates of reaction (from fast to slow) are:Pd(OAc)₂>Pd₂(dba)₃>PdCl₂(CH₃CN)₂. Pd(OAc)₂ and Pd₂(dba)₃ achieved near100% conversion at 5 hours. PdCl₂(CH₃CN)₂ achieved >80% conversion at 5hours and near 100% conversion at 24 hours. Pd₂(dba)₃ (1 mol %)achieved >60% conversion at 24 hours.

The stabilities of the alkyne starting material and the product werealso investigated under reaction conditions. The product appeared stablewhen PdCl₂(MeCN)₂, Pd₂(dba)₃, or Pd(OAc)₂ were used as the catalyst. Thealkyne starting material appeared stable when PdCl₂(MeCN)₂, or Pd₂(dba)₃were used as the catalyst, while degradation of the alkyne startingmaterial was observed when Pd(OAc)₂ was used.

Base Screen

The effect of the base was studied using K₂CO₃, K₃PO₄ and DIPEA with twodifferent metal sources PdCl₂(MeCN)₂ and Pd₂(dba)₃. All experiments wereconducted at 70° C. using acetonitrile as the solvent. The experimentsusing K₂CO₃ and K₃PO₄ as the base showed faster and better conversionsthan DIPEA.

Ligand Screen

Various ligands were screened using the following reaction conditions:Pd(OAc)₂ 5 mol %, 2:1 L:M ratio for monodentate ligands and 1:1 L:Mratio for bidentate ligands, 1.2 eq. alkyne, 1.2 eq K₂CO₃, MeCN (30 rv),70° C. XPhos was used as control reaction. The results are listed in theTable below. The ligands MePhos and cBRIDP were also tested. cBRIDPprovided the highest conversion other than the XPhos control.

Phosphine Conversion Phosphine Conversion BisPhosphine Conversion Ligand(21 h) Ligand (21 h) Ligand (21 h) PCy₃ 50.5 DavePhos 80.7 dppp 30.5PCy₂Ph 39.4 ^(i)Bu Triplecage 4.9 dppe 23.1 P^(i)Pr₃ 20.8 P^(t)Bu₂Cy 2.5dppb 43.5 PCy₂ ^(t)Bu 55.9 P^(t)Bu₃ 61.8 BINAP 52.5 CataCXium A 72.5CataCXium PICy 87.9 DPEPhos 65.6 P(MeOC₆H₄)₃ 53.2 P^(t)Bu₂(PhNMe₂) 52.0dppf 67.0 PPh₂(C₆H₄CO₂H) 10.3 PPh₃ 41.3 dbpf 80.8 PPh₂(C₆H₄SO₃H) 24.2 Noligand 0 XantPhos 58.4 SPhos 65.6 Control reaction 100 N—^(t)Bu₂Pazetine 65.9 JohnPhos 89.2Solvent Screen

Five water immiscible solvents were screened using the followingreaction conditions: Pd₂(dba)₃ 5 mol %, 2:1 L:M ratio for monodentateligands and 1:1 L:M ratio for bidentate ligands, 1.5 eq. alkyne, 1.2 eq.K₂CO₃, solvent (30 rv), 70° C. The results showed that all screenedsolvents worked, and the relative rates of reaction are (from fast toslow): MeCN (control)>^(i)PrOAc>2-MeTHF>EtCN, MEK>>toluene. All screenedsolvents (except toluene) achieved >90% conversion at 5 hours and theperformances were close to MeCN control. Toluene achieved >90%conversion at 24 hours. Water immiscible solvents may be advantageous inwork up to remove Pd residues.

Further studies were carried out to screen for the overall reactionconditions. Three ligands (XPhos, cBRIDP, and CataCXiumPICy), two bases(K₃PO₄ and K₂CO₃), three solvents (MeCN, 2-Me-THF, and toluene),Pd₂(dba)₃ as Pd source with Pd load from 1 to 5 mol % were studied.Based on the results, the condition of Pd₂(dba)₃ (1 mol %), XPhos (2 mol%), K₃PO₄ as the base in MeCN was selected for further studies.

Example 17 Scale Up of Compound C

A 1 L jacketed reaction vessel, equipped with an overhead stirrer,temperature probe, condenser and stirrer was purged with nitrogen for aminimum of 1 hour. Potassium phosphate tribasic (1.2 equ, 42.65 g),4-ethynyl-1-methyl-1H-pyrazole (1.2 equ, 21.34 g) and Compound A (1 wt,1 equ. 50.05 g) were charged to the reaction vessel. Acetonitrile (14volumes, 700 mL) was charged to the reaction vessel. A small amount wasused to rinse out the bottles used for solid charging. Agitation wasstarted. Agitation was set at 470 rpm, as found to be sufficient tosuspend all the solids. Reaction mixture was heated to 40±2° C.(internal temperature).

To a second vessel (100 ml round bottom flask) was charged, in anitrogen inerted glovebox, tris(dibenzylideneacetone)dipalladium(0)(0.01 equ. 1.533 g),2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos) (0.02 equ.1.595 g) and acetonitrile (1 volume, 50 mL). Catalyst/ligand solutionwas stirred at 40° C. for 40 mins.

The first reaction vessel was purged with nitrogen for a further 30minutes before catalyst solution was transferred to the reaction vessel.The catalyst solution was transferred using a needle and syringe tominimise air/moisture exposure. Reaction mixture was heated to 70° C.(internal temperature). Reaction was monitored by HPLC. Reaction wascomplete when sampled at 20.5 hours. Heat was turned off and thereaction mixture was allowed to cool to 25° C. for 2.5 hours.

The suspension was filtered using a glass sinter funnel and the residuewas washed with acetonitrile (1×2 volumes, 100 mL). The wash wascombined with the filtrate (product layer). The filtrate (product layer)was transferred back into the reaction vessel. Reaction vessel wasplaced under a nitrogen purge. 2-Mercaptoethyl ethyl sulfide Silica[PhosphonicS SEM26] (50 wt % wrt Compound A, 25 g) was charged to thevessel. Scavenger was stirred for 6 hours at 28° C. Scavenger wasfiltered off and washed with acetonitrile (1×2 volumes, 100 mL). Thewash was combined with the filtrate (product layer). Solution wastransferred to a clean distillation vessel. Reaction volume was reducedto 5 volumes at atmospheric pressure (jacket temperature of 95-100° C.).Vessel was cooled to 50° C. (internal temperature) over 75 minutes. MTBEwas added slowly (50 mL aliquots) keeping the batch temperature above45° C. MTBE addition was held for 15 minutes once crystallization began.Remaining MTBE added slowly until total of 15 volumes (750 mL) added.Held for 1 hour. Reaction mixture was cooled back to 25° C. over 2 hoursand held overnight. Additional MTBE 5 volumes (250 mL) added over 5minutes. Held for 1 hour. Product was filtered off and washed withacetonitrile (2×2 volumes, 2×100 mL). The wash was combined with themother liquors. Solid was deliquored under vacuum for 15 minutes. Solidwas transferred to crystallization dish and placed in a vacuum oven anddried at 40° C. under vacuum, with an air bleed for 40 hours. Recoveredsolid=52.88 g, Yield=85.8%.

Example 18 Preparation of Compound E

Step 1: i) DIPA (1.3 equ.)/CuI (0.02 equ.)/PdCl₂(PPh₃)₂ (0.01equ.)/TMSacetylene (1.2 equ.)/CH₂Cl₂ (10 vol)/r.t.−40° C./2 hrs; ii)Cool to r.t. and filter; iii) Wash with 2M HCl (1.3 equ.); iv) TreatSiliaMetS Diamine at r.t.; v) Fitler and reduce CH₂Cl₂ to 5 vol. underreduced pressure; vi) Solvent switch to Hexane (5 vol) under reducedpressure; vii) Cool to r.t.; and viii) Filter, wash and dry.

Step 2: Compound F was taken directly into step 2 (crude product);Dissolved in MeOH and treated with K₂CO₃ at r.t.; When the reaction wascomplete the base was filtered off; Solvent evaporated to dryness; Oildissolved in MTBE and passed through a plug of Silica; Filtrateconcentrated to dryness; Product isolated by vacuum distillation (2-3times). Yield=68%.

Example 19 Screening Reaction Conditions for Preparation of Compound F

Step 1 of Example 18 used 0.01 eq. of PdCl₂(PPh₃)₂ and 0.02 eq. of CuI.20 experiments were conducted to investigate whether the amount ofcatalysts can be reduced to avoid or reduce the scavenging needed toremove Palladium/color. The experiments were performed on 2 g scale,with 5 loadings of PdCl₂(PPh₃)₂ (0.001, 0.002, 0.004, 0.0055, and 0.01eq.) and 4 loadings of CuI (0.001, 0.005, 0.0125, and 0.02 eq.). IPC ofthe reaction mixture at different time points: 4 min, 30 min, 1 hr, 2hrs, 4 hrs, 6 hrs, 8 hrs and 24 hrs. Samples analyzed by HPLC todetermine evolution of conversion and % area of SM and impurities(particularly the homocoupling diyne side-product

FIG. 34A shows the reaction conversion at 2 hours (% area); FIG. 34Bshows the reaction conversion at 24 hours (% area); FIG. 34C shows theproduct/diyne ratio (% area) at 24 hours; and FIG. 34D shows the diynecontent at 24 hours (% area). The results show that the reactionconversion is affected by both Pd and Cu content, but the diyneformation is primarily affected by Pd content. Copper has a biggerimpact when the Pd content is low. Reaction can reach completion withlower levels of Pd. Reducing the amount of Pd can reduce the formationof the diyne impurity. Based on the results, 0.003 eq. of Pd catalystand 0.0175 eq. of CuI may be an optimal condition.

An exemplary study using 0.003 eq. of Pd catalyst and 0.0175 eq. of CuIwas conducted as follows. 4-iodo-1-methylpyrazole (250 g, 1190 mmol),copper iodide (3.99 g, 20.82 mmol, 0.0175 eq.), andbis(triphenylphosphine)palladium(II) chloride (2.53 g, 3.57 mmol, 0.003eq) were charged to a reactor at r.t under nitrogen. DIPA (219 mL, 1547mmol) was charged to the reactor under nitrogen at 22° C. via syringe ina single portion. ACS grade DCM (2500 mL) was charged. The reactor wasflushed with nitrogen for ≥5 minutes and the temperature was set to 15°C. Trimethylsilylacetylene (205 mL, 1428 mmol) was charged via syringepump over 30 minutes (or 82 mL/hr or 1.4 ml/min) at 15° C. 10 mL oftoluene was added in the flask at the end of the addition to rinse thevessel. The reaction was monitored by HPLC after 24 hours (reactioncompleted between 8-24 hours).

HCl (1785 mL, 3570 mmol) was charged at 15±3° C. and the mixture wasstirred vigorously for ≥15 minutes. The mixture was then allowed tosettle and the layers were separated over ≥10 minutes. The upper layerwas clear yellow and the lower was clear orange. The layers were split,and the upper layer was discarded. 10 volumes (2500 mL) of 20% wt sodiumthiosulfate was charged to the lower layer to scavenge palladium andcopper. The mixture was stirred for overnight under nitrogen at r.t. Thephases were allowed to separate over ≥10 minutes. The layers wereseparated.

The organic layer was transferred back into the cleaned reactor, anddistilled down to 5 volumes (1250 mL). 5 volumes of hexanes (1250 mL)was added and distilled down to 5 volumes. 5 volumes of hexanes (1250mL) was added again and distilled down to 5 volumes. Another 5 volumesof hexanes (1250 mL) was added and heated to reflux during 1 hour, andthen cooled down to r.t. The mixture was filtered with a Whatman filterpaper to remove the alkyne dimer side-product. The mother liquor thecrystals formed in it were transferred back to a clean reactor. Thesolvent was distilled down to 5 volumes (1250 mL), cooled down to 24° C.over an hour, then cooled down to 0° C., and aged for an hour. Thecrystal product was filtered, washed with 2×1 volume of cold hexanes,dried, and collected.

Example 20 Screening Reaction Conditions for Preparation of Compound E

The deprotection of Compound F in biphasic solvent system (mixture ofaqueous base solution and a water immiscible solvent) was investigated.The results are summarized in the table below. The experiments wereperformed on 100 mg scale. Using heat and a phase transfer catalyst didnot improve the yield.

MTBE DCM Ethyl Acetate 2 hrs 15 hrs 24 hrs 2 hrs 15 hrs 24 hrs 2 hrs 15hrs 24 hrs KOH   92%   100% X 17.87% 93.20% 98.39% 0% 0% 0% NaOH 6.90%67.90% 85.28% 4.28% 18.90% 26.70% 0% 0% 0% NaHCO₃   0%  2.64% 2.74%0.00%    0%    0% 0% 0% 0% K₃PO₄ 5.78% 96.46% 100.00% 5.21% 37.79%54.62% 3.81%   4.22%   4.96%   K₂CO₃ 9.98% 62.22% 78.96% 1.04% 10.48%14.54% 1.05%   2.88%   3.68%  

Example 21 Scale Up of Compound E

Step 1:

A clean and dry 500 L glass-lined reactor A was evacuated to −0.08-−0.05MPa and filled with nitrogen to normal pressure. It was repeated for 3times. The reactors were sampled for oxygen content to ensure it was≤3%.

Diisopropylamine (106.0 kg) and 1-methyl-4-iodo-1H-pyrazole (24.3 kg,23.6 kg corrected) were added to the reactor A at 15-25° C. Cuprousiodide (0.37 kg) was added to reactor A under the protection of nitrogenat 15-25° C. Bis(triphenylphosphine)palladium(II) chloride (1.09 kg) wasadded to the mixture under the protection of nitrogen at 15-25° C. Themixture was stirred for 20-30 min. Trimethylsilylacetylene (22.2 kg,14.0 kg corrected) was added to the mixture in portions with 4-5 kg andan interval of 20-30 min for each portion at 15-30° C. The mixture wasallowed to react at 20-30° C. After 2 h, the mixture was sampled every1-2 h for purity analysis by HPLC until area % of1-methyl-4-iodo-1H-pyrazole was ≤0.5%.

After reaction completion, the mixture was filtered with a stainlesssteel centrifuge. The filter cake was rinsed twice with methyltert-butyl ether (9.2 kg×2). The filtrate was transferred to reactor Aand concentrated under reduced pressure (P≤−0.08 MPa) at T≤45° C. until40-60 was left. Methyl tert-butyl ether (92.5 kg) was added to themixture and concentration was continued until 40-60 L was left. Methyltert-butyl ether (92.2 kg) was added to concentrated mixture and themixture was sampled for diisopropylamine residual analysis to ensure itwas ≤1%. Active carbon (4.9 kg) was added to the mixture at 15-25° C.and the mixture was maintained for 6-8 h under stirring. The mixture wasfiltered with stainless steel nutsche filter at 15-25° C. The filtercake was rinsed twice with methyl tert-butyl ether (9.2 kg×2). Asolution of citric acid monohydrate (6.1 kg) in purified water (121.6kg) was added to the filtrate at 15-25° C. The mixture was stirred for20-30 min and settled for 20-30 min before separation. The emulsionlayer was separated to aqueous phase. The aqueous phase was sampled forpH analysis and wt % analysis to ensure pH was ≤7. Active carbon (4.9kg) was added to the mixture at 15-25° C. and the mixture was maintainedfor 6-8 h under stirring. The mixture was filtered with stainless steelnutsche filter at 15-25° C. The filter cake was rinsed twice with methyltert-butyl ether (9.2 kg×2). The filtrate was checked to ensure it wasyellow solution.

The filtrate was transferred to reactor B and concentrated under reducedpressure (P≤−0.08 MPa) at T≤35° C. until 30-40 L was left. Anhydrousethanol (96.3 kg) was added to the mixture and concentration wascontinued at T≤45° C. until 30-40 L was left. The mixture was sampledfor methyl tert-butyl ether residual analysis to ensure it was ≤0.5%.The mixture was cooled to 15-25° C. Purified water (121.3 kg) was addedto the mixture through peristaltic pump at 15-25° C. at a reference rateof 25-50 kg/h. Brown yellow solid precipitated.

The mixture was allowed to crystallize at 15-25° C. After 2 h, themixture was sampled every 1-2 h for mother liquor wt % analysis until itwas ≤0.5% or the difference between the two samples was ≤0.3%. Themixture was filtered with a stainless steel nutsche filter. The filtercake was rinsed twice with purified water (12.1 kg×2). The filter cakewas swept in the stainless steel nutsche filter. After 12 h, the solidwas sampled every 6-8 h for ethanol residual until it was ≤1%. Theproduct was packaged in one plastic bag. Product weight=12.0 kg,Yield=52.3%, Purity (HPLC)=98.9%.

Step 2:

A clean and dry 1000 L glass-lined reactor A and a glass-lined reactor500 L reactor B were evacuated to −0.08-−0.05 MPa and filled withnitrogen to normal pressure. It was repeated for 3 times. The reactorswere sampled for oxygen content to ensure it was no more than 3%.

Methyl tert-butyl ether (88.8 kg) and Compound F (12.0 kg crude fromStep 1, 10.5 kg adjusted) were added to the reactor A at 15-25° C. Thestirrer was started. Then a solution of potassium hydroxide (12.0 kg) inpurified water (108.0 kg) was added to the mixture at 15-25° C. Themixture was allowed to react at 15-25° C. After 4 h, the mixture wassampled every 2-4 h for purity analysis by HPLC until area % of CompoundF was no more than 1.0%.

Active carbon (1.2 kg) was added to the mixture at 15-25° C. The mixturewas stirred for 2-3 h. The mixture was filtered with a stainless steelnutsche filter at 15-25° C. The filter cake was rinsed twice with methyltert-butyl ether (22.2 kg×2). The filtrate was stirred for 20-30 min andsettled for 20-30 min at 15-25° C. before separation. The mother liquorof step 1 (354.9 kg) and methyl tert-butyl ether (205.6 kg) were addedto reactor B. The mixture was stirred for 30-40 min and settled for20-30 min at 15-25° C. before separation. Active carbon (1.2 kg) wasadded to the organic phase at 15-25° C. The mixture was stirred for 2-3h. The mixture was filtered with a stainless steel nutsche filter at15-25° C. The filter cake was rinsed twice with methyl tert-butyl ether(22.2 kg×2). The organic phases and the filtrate were combined intoreactor B and concentrated under reduced pressure at T≤25° C. until160-190 L was left. The mixture was transferred to 200 L iron drum.Methyl tert-butyl ether (24.2 kg) was added to reactor B to rinsereactor wall and the rinsing liquor was combined with the mixture.

A clean and dry 80 L glass reactor C was evacuated to −0.08-−0.05 MPaand filled with nitrogen to normal pressure. It was repeated for 3times. The reactor was sampled for oxygen content to ensure it was ≤3%.The mixture was transferred to reactor C in portions and concentratedunder reduced pressure at T≤25° C. (until 25 L was left.

n-Heptane (16.3 kg) was added to the mixture at 15-25° C. and themixture was sampled for methyl tert-butyl ether residual analysis. Themixture was heated to 55-60° C. under stirring. Then the mixture wascooled to −10-0° C. at a reference rate of 10-15° C./h. The mixture wasstirred at −10-0° C. for crystallization. After 1-2 h, the mixture wassampled every 2-3 h for wt % analysis of mother liquor until it was ≤3wt %. The mixture was filtered with a filter flask and the filter cakewas rinsed twice with pre-cooled n-heptane (8.1 kg total, −10-0° C.).The filter cake and methyl tert-butyl ether (8.9 kg) were added to 20 Lflask at 15-25° C. The mixture was stirred until all solid was dissolvedcompletely by visual check.

The mixture was transferred to reactor C and methyl tert-butyl ether(35.5 kg) was added to the mixture through in-line filter at 15-25° C.The mixture was concentrated under reduced pressure at T≤25° C. until10-15 L was left. Methyl tert-butyl ether (˜26.6 kg×2) was added to themixture and the concentration was continued. Methyl tert-butyl ether(26.7 kg) was added to the mixture and the mixture was sampled for KFanalysis to ensure it was ≤0.4%. The mixture was concentrated underreduced pressure at T≤25° C. until no distillate was observed (˜10 L wasleft). The mixture was transferred to 10 L flask E and evacuated underreduced pressure with oil pump until solid was formed. The product washeated to 25-30° C. and then transferred to six liquid pails. Innerpackage: bagged with pharmaceutical grade HDPE liquid pail. Outerpackage: pharmaceutical grade LDPE bag with food grade security seal.Product weight 6.1 kg, Yield=94.4%, Purity (HPLC)=98.9%.

Example 22 Preparation of Compound E

Step 1 was conducted according to the procedure as Step 1 of Example 18.

Step 2: At the end of the reaction the organic layer was separated off.The solvent was distilled off under vacuum followed by Compound E.Yield=85-88% as a colorless crystalline solid.

When Compound F crude product was telescoped into Step 2 without furtherpurification, the overall yield of the two step was about 57%. The phasesplit was difficult because both layers had dark brown and solidmaterial was present. A carbon treatment may be necessary to remove someof the color. The distilled Compound E was provided as a yellow oil.

Alternatively, Compound F was recrystallized after Step 1 before beingused in Step 2. The recrystallization yield was about 71-78%, and about25% of Compound F was lost to the mother liquid. The overall yield ofthe two step was about 85-88%. Only a phase split and singledistillation was required to afford Compound E as a colorless crystal.The brine wash and carbon treatment from the telescope approach can beavoided.

Example 23 Preparation of Compound M

Recrystallization from MeOH led to relatively high loss due to moderatesolubility.

Re-slurry with 1-propanol was less effective on larger scale or whenclumps were present.

An exemplary study where 1-butanol was used for crystallization wascarried out as follow. The solvent of 1-BuOH is convenient as it is alsothe solvent for the next reaction.

MeCN (15 volumes) and NaOH (2 equivalents) were charged to anappropriately sized jacketed reactor. Mixing was started and the jacketwas set to maintain an internal temperature of 15° C. Ethyl cyanoacetatewas charged to the reactor over a period of 20 to 30 minutes, adjustingthe jacket to maintain temperature as necessary. Once the charge wascomplete, the jacket was set to maintain 20° C. and stirring wascontinued for 2 h. The exotherm from the deprotonation was mild and waseasy to control even at larger scales. After two hours, the jacket wasset to maintain an internal temperature of 10° C. A solution of2-chloroethyl chloroformate (1 equivalent) in MeCN (2 volumes) wascharged to the reactor over the course of at least 1 hour. The jacketwas adjusted as necessary to maintain the internal temperature at 10-15°C. Once the charge was completed the solution was refluxed for 1 hour.Following reflux, the reactor contents were cooled to room temperatureand the NaCl generated was removed by filtration. The solution can beheld overnight at this point if required.

The solution was charged back to the cleaned reactor and distilled undervacuum to remove MeCN until 5 volumes remained. Once at five volumes,the vacuum was broke and 5 volumes of 1-BuOH was charged to the reactor.The distillation was restarted and continued to 5 volumes. During thecourse of this distillation the material started to precipitate. Once atfive volumes, the vacuum was broke and a sample of the mother liquor wastaken for analysis by NMR or GC. A passing IPC (in-process control) atthis point required <5 mol % MeCN in 1-BuOH. If the IPC failed,distillation was continued and re-sampled at a later time point, the IPCwould eventually pass. Once a passing IPC was obtained, the slurry wascooled to room temperature and held overnight if necessary.

The slurry was filtered using a Buchner funnel and the cake was slurrywashed once with 1.33 volumes 1-BuOH and displacement washed once with1.33 volumes of 1-BuOH. The cake was dried in a vacuum oven untilreaching constant weight. The typical yield of this step is 75-80% andthe reaction had been carried out on 225 g scale.

Example 24 Preparation of Compound J

An exemplary study where 1-butanol was used as solvent was carried outas follow.

Compound M and 1-BuOH (6.5 volumes) were charged to an appropriatelysized jacketed reactor. Mixing was started and the jacket was set tomaintain an internal temperature of 25° C. Aqueous ammonia (1.1equivalents) was charged to the reactor. There was an initial endothermfrom the dissolution of the ammonia and then a very mild exotherm as thereaction begins. An IPC sample should be taken following a minimum of1.5 hours and the passing criteria is <1 area % remaining startingmaterial. Once a passing IPC was obtained, hydrazine monohydrate (1.5equivalents) was charged to the reactor and the jacket was set tomaintain an internal temperature of 70° C. After 2 hours an IPC samplecan be taken and a passing test has <1 area % remaining intermediate(Compound L). Once this passing IPC occurred, the jacket was set tomaintain an internal temperature of 105-110° C. (reflux). The reactionwas then allowed to proceed for about 40 hours.

The solution was then cooled to 80° C. and an IPC sample was taken. Thereaction was then re-heated to reflux at this time. The passing criteriawas <1 area % remaining intermediate (Compound K). Once this passing IPCwas obtained, the solution was cooled over 2 hours to 20° C. and theslurry was held for 2 h once that temperature was reached. The slurrywas filtered using a Buchner funnel and the cake slurry was washed twicewith 2 volumes of 1-BuOH and displacement washed once with 1 volume of1-BuOH. The cake was dried in a vacuum oven to constant weight. Thisprocedure had been carried out on 200 g scale.

Example 25 Preparation of Compound G

In an exemplary study, 3 volumes of AcOH were used. AcOH is distilledoff under mild vacuum at elevated temperature to keep in solution. Oncedistillation ended, LiOH solution was added directly. Overall purity wasabout 95%.

Another exemplary study was carried out as follow: Compound J and AcOH(7.5 volumes) were charged to an appropriately sized jacketed reactor.Mixing was started and the jacket was set to maintain an internaltemperature of 25° C. Tetramethoxypropane (1.01 equivalents) was chargedto the reactor and the jacket was set to maintain an internaltemperature of 95° C. Once at temperature, the reaction continued mixingfor 1.5 hours and then an IPC sample was taken. The passing criteria forthis IPC was <1 area % remaining Compound J. If necessary, a furtheraddition of 0.05 equivalents of tetramethoxypropane may be added to pushthe reaction to completion. Once a passing IPC was obtained, thesolution was concentrated to 3 volumes by vacuum distillationmaintaining a solution temperature >60° C. to prevent prematureprecipitation. Once the target volume was reached the jacket was set tomaintain an internal temperature of 50° C. A 4 M solution of NaOH wasthen charged to the reactor via cannula to neutralize the remainingAcOH. This typically required approximately 10 volumes of the basesolution. The neutralization was monitored by pH probe. Solids began toprecipitate during the course of the charge.

Once neutralized, the slurry was cooled to 20° C. and held at thattemperature for 1 hour prior to isolation via Buchner funnel. The cakewas washed twice with 2 volumes of water and once with 2 volumes ofMeOH. The solids were then dried to constant weight in a vacuum oven toprovide Compound H. This procedure had been performed on 110 g scale toproduce a granular light brown solid.

Compound H and MeOH (3.6 volumes) were charged to an appropriately sizedjacketed reactor. Mixing was started and the jacket was set to maintainan internal temperature of 25° C. A solution of LiOH (4.4 equivalents)in water (12 volumes) was charged to the reactor slowly over severalminutes. There was a small exotherm upon charging, but it was small andeasily managed. Once the charge was complete, the slurry was heated to50° C. for a minimum of 2 hours. After this time an IPC sample was takenwith a targeted passing specification of <1 area % Compound H. Once apassing IPC was obtained, the jacket was set to maintain the slurry at10° C. pH was adjusted to 4 using 6 M and 1 M HCl transferred viacannula. The 6 M HCl was used for the initial adjustment and the 1 M HClwas used for the end of the titration. The slurry was held overnight atthis point if necessary.

The slurry was filtered with a Buchner funnel and the cake was washedtwice with 3 volumes of water and once with 3 volumes of MeOH. The cakewas then dried to constant weight in a vacuum oven to provide CompoundG. This procedure had been performed on 122 g scale to produce a paleyellow solid.

In an alternative example, the reaction was carried out using HCl as theacid instead of AcOH.

The overall average yield in three steps (preparation of Compound M,preparation of Compound J, and preparation of Compound G) was about40-45%, and the product was obtained in off white color.

While exemplary embodiments of the present disclosure have been shownand described herein, it will be obvious to those skilled in the artthat such embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the disclosure. It should beunderstood that various alternatives to the embodiments described hereincan be employed in practicing the subject matter of the disclosure. Allsuch equivalents are considered to be within the scope of the claimedsubject matter and are encompassed by the appended claims.

What is claimed is:
 1. A solid form of a compound of Formula (I), of asolvate of a compound of Formula (I), or of a cocrystal of a compound ofFormula (I):

wherein the solid form is: Form 1 of a compound of formula (I), havingan X-ray powder diffraction (XRPD) pattern comprising peaks at 16.8,23.6, and 25.6 degrees 2θ, plus or minus 0.2; Form 2 of a 1-propanol,acetone, chloroform, or dichloromethane solvate of a compound of formula(I), having an XRPD pattern comprising peaks at 13.6, 14.9, and 21.0degrees 2θ, plus or minus 0.2; Form 3 of a MEK, 1-propanol, acetone,dimethyl sulfoxide, tert-butylmethyl ether, 2-methyl-1-propanol, or2-methyl THF solvate of a compound of formula (I), having an XRPDpattern comprising peaks at 17.9, 20.6, and 25.8 degrees 2θ, plus orminus 0.2; Form 4 of a 2-propanol, 1-propanol, dimethyl sulfoxide,2-methyl-1-propanol, 1,4-dioxane, chloroform, THF, 2-methoxyethanol, oracetone solvate of a compound of formula (I), having an XRPD patterncomprising peaks at 7.4, 18.0, and 20.7 degrees 2θ, plus or minus 0.2;Form 5 of an anisole solvate of a compound of formula (I), having anXRPD pattern comprising peaks at 21.0, 22.1, and 25.2 degrees 2θ, plusor minus 0.2; Form 6 of a hydrate of a compound of formula (I), havingan XRPD pattern comprising peaks at 4.8, 19.9, and 26.7 degrees 2θ, plusor minus 0.2; Form 7 of a MEK, 1-propanol, acetone, or tert-butylmethylether solvate of a compound of formula (I), having an XRPD patterncomprising peaks at 7.5, 12.3, and 20.7 degrees 2θ, plus or minus 0.2;Form 8 of a compound of formula (I), having an XRPD pattern comprisingpeaks at 18.8,20.8, and 24.5 degrees 2θ, plus or minus 0.2; Form P1C3 ofa cocrystal of a compound of Formula (I) and L-tartaric acid, having anXRPD pattern comprising peaks at 11.2, 17.4, and 17.7 degrees 2θ, plusor minus 0.2; Form P1C9 of a cocrystal of a compound of Formula (I) andsalicylic acid, having an XRPD pattern comprising peaks at 6.9, 10.1,and 12.0 degrees 2θ, plus or minus 0.2; or Form P2C9 of a cocrystal of acompound of Formula (I) and salicylic acid, having an XRPD patterncomprising peaks at 11.4, 13.4, and 24.0 degrees 2θ, plus or minus 0.2.2. The solid form of claim 1, wherein the solid form is Form 1 of acompound of formula (I), having an X-ray powder diffraction (XRPD)pattern comprising peaks at 16.8, 23.6, and 25.6 degrees 2θ, plus orminus 0.2.
 3. The solid form of claim 2, having an XRPD pattern furthercomprising at least one peak selected from 14.6 and 21.2 degrees 2θ,plus or minus 0.2.
 4. The solid form of claim 2, having an XRPD patternsubstantially as shown in FIG.
 1. 5. The solid form of claim 2, whichexhibits an endothermic event, as characterized by DSC, with an onsettemperature at about 255° C. and/or a peak temperature at about 257° C.6. The solid form of claim 2, which exhibits a weight loss of about 0.6%upon heating from about 230° C. to about 310° C.
 7. The solid form ofclaim 2, which exhibits a weight increase of about 0.5% when subjectedto an increase in relative humidity from about 0 to about 90% relativehumidity.
 8. The solid form of claim 2, having approximately unit celldimensions of: a=11.1 Å, b=12.8 Å, c=16.1 Å, α=90°, β=90°, and γ=90°. 9.The solid form of claim 2, which is anhydrous.
 10. The solid form ofclaim 2, which is non-hygroscopic.
 11. The solid form of claim 1,wherein the solid form is Form 2 of a 1-propanol, acetone, chloroform,or dichloromethane solvate of a compound of formula (I), having an XRPDpattern comprising peaks at 13.6, 14.9, and 21.0 degrees 2θ, plus orminus 0.2.
 12. The solid form of claim 11, having an XRPD patternfurther comprising at least one peak selected from 7.4 and 16.7 degrees2θ, plus or minus 0.2.
 13. The solid form of claim 11, having an XRPDpattern substantially as shown in FIG.
 4. 14. The solid form of claim11, having approximately unit cell dimensions of: a=8.7 Å, b=13.2 Å,c=26.0 Å, α=90°, β=90°, and γ=90°.
 15. The solid form of claim 1,wherein the solid form is Form 3 of a MEK, 1-propanol, acetone, dimethylsulfoxide, tert-butylmethyl ether, 2-methyl-1-propanol, or 2-methyl THFsolvate of a compound of formula (I), having an XRPD pattern comprisingpeaks at 17.9, 20.6, and 25.8 degrees 2θ, plus or minus 0.2.
 16. Thesolid form of claim 15, having an XRPD pattern further comprising atleast one peak selected from 11.7 and 23.5 degrees 2θ, plus or minus0.2.
 17. The solid form of claim 15, having an XRPD patternsubstantially as shown in FIG.
 9. 18. The solid form of claim 1, whereinthe solid form is Form 4 of a 2-propanol, 1-propanol, dimethylsulfoxide, 2-methyl-1-propanol, 1,4-dioxane, chloroform, THF,2-methoxyethanol, or acetone solvate of a compound of formula (I),having an XRPD pattern comprising peaks at 7.4, 18.0, and 20.7 degrees2θ, plus or minus 0.2.
 19. The solid form of claim 18, having an XRPDpattern further comprising at least one peak selected from 11.9 and 13.6degrees 2θ, plus or minus 0.2.
 20. The solid form of claim 18, having anXRPD pattern substantially as shown in FIG.
 11. 21. The solid form ofclaim 1, wherein the solid form is Form 5 of an anisole solvate of acompound of formula (I), having an XRPD pattern comprising peaks at21.0, 22.1, and 25.2 degrees 2θ, plus or minus 0.2.
 22. The solid formof claim 21, having an XRPD pattern further comprising at least one peakselected from 14.5 and 19.2 degrees 2θ, plus or minus 0.2.
 23. The solidform of claim 21, having an XRPD pattern substantially as shown in FIG.13.
 24. The solid form of claim 1, wherein the solid form is Form 6 of ahydrate of a compound of formula (I), having an XRPD pattern comprisingpeaks at 4.8, 19.9, and 26.7 degrees 2θ, plus or minus 0.2.
 25. Thesolid form of claim 24, having an XRPD pattern further comprising atleast one peak selected from 11.9 and 24.8 degrees 2θ, plus or minus0.2.
 26. The solid form of claim 24, having an XRPD patternsubstantially as shown in FIG.
 15. 27. The solid form of claim 1,wherein the solid form is Form 7 of a MEK, 1-propanol, acetone, ortert-butylmethyl ether solvate of a compound of formula (I), having anXRPD pattern comprising peaks at 7.5, 12.3, and 20.7 degrees 2θ, plus orminus 0.2.
 28. The solid form of claim 27, having an XRPD patternfurther comprising at least one peak selected from 13.7 and 17.2 degrees2θ, plus or minus 0.2.
 29. The solid form of claim 27, having an XRPDpattern substantially as shown in FIG.
 18. 30. The solid form of claim1, wherein the solid form is Form 8 of a compound of formula (I), havingan XRPD pattern comprising peaks at 18.8, 20.8, and 24.5 degrees 2θ,plus or minus 0.2.
 31. The solid form of claim 30, having an XRPDpattern further comprising at least one peak selected from 16.0 and 17.9degrees 2θ, plus or minus 0.2.
 32. The solid form of claim 30, having anXRPD pattern substantially as shown in FIG.
 20. 33. The solid form ofclaim 1, wherein the solid form is Form P1C3 of a cocrystal of acompound of Formula (I) and L-tartaric acid, having an XRPD patterncomprising peaks at 11.2, 17.4, and 17.7 degrees 2θ, plus or minus 0.2.34. The solid form of claim 33, having an XRPD pattern furthercomprising at least one peak selected from 21.2 and 22.5 degrees 2θ,plus or minus 0.2.
 35. The solid form of claim 33, having an XRPDpattern substantially as shown in FIG.
 22. 36. The solid form of claim1, wherein the solid form is Form P1C9 of a cocrystal of a compound ofFormula (I) and salicylic acid, having an XRPD pattern comprising peaksat 6.9, 10.1, and 12.0 degrees 2θ, plus or minus 0.2.
 37. The solid formof claim 36, having an XRPD pattern further comprising at least one peakselected from 17.8 and 20.0 degrees 2θ, plus or minus 0.2.
 38. The solidform of claim 36, having an XRPD pattern substantially as shown in FIG.24.
 39. The solid form of claim 1, wherein the solid form is Form P2C9of a cocrystal of a compound of Formula (I) and salicylic acid, havingan XRPD pattern comprising peaks at 11.4, 13.4, and 24.0 degrees 2θ,plus or minus 0.2.
 40. The solid form of claim 39, having an XRPDpattern further comprising at least one peak selected from 25.1 and 26.9degrees 2θ, plus or minus 0.2.
 41. The solid form of claim 39, having anXRPD pattern substantially as shown in FIG.
 26. 42. A pharmaceuticalcomposition comprising a solid form of claim 1 and a pharmaceuticalacceptable excipient thereof.
 43. A pharmaceutical compositionconsisting essentially of the solid form of claim 1.